Heterobicyclic sphingosine 1-phosphate analogs

ABSTRACT

Compounds that have agonist activity at one or more of the S1P receptors are provided. The compounds are sphingosine analogs that, after phosphorylation, can behave as agonists at S1P receptors.

CLAIM OF PRIORITY

This application claims priority to U.S. Provisional Patent ApplicationNo. 61/109,720, filed on Oct. 30, 2008, which is incorporated byreference in its entirety.

BACKGROUND

Sphingosine 1-phosphate (S1P) is a lysophospholipid mediator that evokesa variety of cellular responses by stimulation of five members of theendothelial cell differentiation gene (EDG) receptor family. The EDGreceptors are G-protein coupled receptors (GPCRs) and on stimulationpropagate second messenger signals via activation of heterotrimericG-protein alpha (G_(α)) subunits and beta-gamma (G_(βγ)) dimers.Ultimately, this S1P-driven signaling results in cell survival,increased cell migration and, often, mitogenesis. The recent developmentof agonists targeting S1P receptors has provided insight regarding therole of this is signaling system in physiologic homeostasis. Forexample, the immunomodulator, FTY720(2-amino-2-[2-(4-octylphenyl)ethyl]propane 1,3-diol), that followingphosphorylation, is an agonist at 4 of 5 S1P receptors, revealed thatenhancing S1P tone influences lymphocyte trafficking. Further, S1P type1 receptor (S1P₁) antagonists cause leakage of the lung capillaryendothelium, which suggests that S1P may be involved in maintaining theintegrity of the endothelial barrier in some tissue beds.

S1P has been demonstrated to induce many cellular processes, includingthose that result in platelet aggregation, cell proliferation, cellmorphology, tumor-cell invasion, endothelial cell chemotaxis andangiogenesis. For these reasons, S1P receptors are good targets fortherapeutic applications such as wound healing and tumor growthinhibition. Sphingosine-1-phosphate signals cells in part via a set of Gprotein-coupled receptors named S1P₁, S1P₂, S1P₃, S1P₄, and S1P₅(formerly EDG1, EDG5, EDG6 and EDG8). The EDG receptors are G-proteincoupled receptors (GPCRs) and on stimulation propagate second messengersignals via activation of heterotrimeric G-protein alpha (G_(α))subunits and beta-gamma (G_(βγ)) dimers. These receptors share 50-55%amino acid sequence identity and cluster with three other receptors(LPA₁, LPA₂, and LPA₃ (formerly EDG2, EDG4 and EDG7) for thestructurally related lysophosphatidic acid (LPA).

A conformational shift is induced in the G-Protein Coupled Receptor(GPCR) when the ligand binds to that receptor, causing GDP to bereplaced by GTP on the α-subunit of the associated G-proteins andsubsequent release of the G-proteins into the cytoplasm. The α-subunitthen dissociates from the βγ-subunit and each subunit can then associatewith effector proteins, which activate second messengers leading to acellular response. Eventually the GTP on the G-proteins is hydrolyzed toGDP and the subunits of the G-proteins reassociate with each other andthen with the receptor. Amplification plays a major role in the generalGPCR pathway. The binding of one ligand to one receptor leads to theactivation of many G-proteins, each capable of associating with manyeffector proteins leading to an amplified cellular response.

S1P receptors make good drug targets because individual receptors areboth tissue and response specific. Tissue specificity of the S1Preceptors is desirable because development of an agonist or antagonistselective for one receptor localizes the cellular response to tissuescontaining that receptor, limiting unwanted side effects. Responsespecificity of the S1P receptors is also of importance because it allowsfor the development of agonists or antagonists that initiate or suppresscertain cellular responses without affecting other responses. Forexample, the response specificity of the S1P receptors could allow foran S1P mimetic that initiates platelet aggregation without affectingcell morphology.

Sphingosine-1-phosphate is formed as a metabolite of sphingosine in itsreaction with sphingosine kinase and is stored in abundance in theaggregates of platelets where high levels of sphingosine kinase existand sphingosine lyase is lacking. S1P is released during plateletaggregation, accumulates in serum, and is also found in malignantascites. Reversible biodegradation of S1P most likely proceeds viahydrolysis by ectophosphohydrolases, specifically the sphingosine1-phosphate phosphohydrolases. Irreversible degradation of S1P iscatalyzed by S1P lyase yielding ethanolamine phosphate and hexadecenal.

A class of S1P agonist compounds are described in provisional U.S.Application No. 60/956,111, filed Aug. 15, 2007, and PCT/US2008/073378,filed Aug. 15, 2008, each of which is incorporated by reference in itsentirety.

SUMMARY

Currently, there is a need for novel, potent, and selective agents thatare agonists of the S1P receptor having enhanced potency, selectivity,and oral bioavailability. In addition, there is a need in the art foridentification of, as well as the synthesis and use of, such compounds.

In one aspect, a compound of formula (I):

in which A¹ can be —C(X¹)═, —N═, —O—, —S—, or a bond; A² can be —C(X²)═,—N═, —O—, —S—, or a bond; A³ can be —C(X³)(X^(3′))—, —C(X³)═, —NX³—,—N═, —O—, or —S—; A⁴ is —C(X⁴)(X^(4′))—, —C(X⁴)═, —NX⁴—, —N═, —O—, or abond; A⁵ can be —C(X⁵)(X^(5′))—, —C(X⁵)═, —NX⁵—, —N═, —O—, or —S—; andA⁶ can be —C(X⁶)═, —N═, —O—, —S—, or a bond; provided that A¹, A², A³,A⁴, A⁵ and A⁶ are not simultaneously —C(X¹)═, —C(X²)═, —C(X³)═, —C(X⁴)═,—C(X⁵)═, and —C(X⁶)═ respectively, and provided that the bicyclic ringincludes 0-3 heteroatoms; and further provided that no more than one ofA¹, A², and A⁶ is a bond.

Each of X¹, X², X³, X^(3′), X⁴, X^(4′), X⁵, X^(5′), and X⁶,independently, can be hydrogen, halo, hydroxy, nitro, cyano, alkyl,haloalkyl, cycloalkyl, halocycloalkyl, alkoxy, haloalkoxy, cycloalkoxy,halocycloalkoxy, acyl, aminoacyl, —NR^(f)R^(g), —N(R^(f))SO₂R^(g),—SO₂R^(f), —SO₂NR^(f)R^(g), —CO₂R^(f), trialkylamino, aryl, orheteroaryl.

Y can be —OR^(f), —(CR^(f)R^(g))OR^(f), —(CR^(f)R^(g))₂OR^(f),—O—P(O)(OR^(f))OR^(g), —OC(O)R^(c), —C(O)OR^(c),—(CR^(f)R^(g))—P(O)(OR^(f))OR^(g), —(C(OH)R^(f))—P(O)(OR^(f))OR^(g),—S—P(O)(OR^(f))OR^(g), tetrazole, —SO₂NHR^(f), —SO₃, —CONHR^(f),—Si(OH)₂, or —B(OH)₂.

W can be —CR^(f)R^(g)—, —NR^(f)—, —O—, —S—, —SO—, or —SO₂—.

Cy can be cycloalkyl, cycloalkenyl, heterocyclyl, aryl, or heteroaryl;wherein Cy is optionally substituted by 1-6 substituents selected fromthe group consisting of hydrogen, halo, hydroxy, nitro, cyano,—NR^(f)R^(g), alkyl, haloalkyl, cycloalkyl, cycloalkenyl,cycloalkylalkyl, cycloalkenylalkyl, heterocyclylalkyl, arylalkyl,heteroarylalkyl, alkoxy, haloalkoxy, cycloalkylalkoxy,cycloalkenylalkoxy, heterocyclylalkoxy, aryloxy, arylalkoxy,heteroaryloxy, heteroarylalkoxy, acyl, cycloalkylacyl, cycloalkenylacyl,heterocyclylacyl, arylacyl, heteroarylacyl, thioalkyl, alkenyl, alkynyl,cycloalkenyl, heterocyclyl, aryl, and heteroaryl.

L¹ can be —CH₂—, —CHF—, or —CF₂—.

Z⁴ can be hydrogen, halo, alkyl, haloalkyl, alkenyl, haloalkenyl,alkynyl, or —OR^(f); or Z⁴ can be —CH₂— bound to the carbon atom towhich Y is bound; or L¹, Z⁴, Y, and the atoms to which they are boundcan form a 4-7 membered cycloalkyl group or a 4-7 membered heterocyclylgroup having 1 or 2 heteroatoms selected from O and N.

R^(a) can be hydrogen, alkyl, haloalkyl, alkenyl, alkynyl, cycloalkyl,cycloalkenyl, aryl, heteroaryl, or heterocyclyl; wherein each of alkyl,cycloalkyl, cycloalkenyl, aryl, heteroaryl and to heterocycle areoptionally substituted with 1 to 5 substituents independently selectedfrom the group consisting of halo, oxo, —CN, —CHO, —CF₃, —OH, —NO₂,alkyl, —OCF₃, alkoxy, cycloalkoxy, cycloalkenoxy, amino, alkylamino,dialkylamino, acylamino, aminoacyl, alkylsulfonyl, alkylaminosulfonyl,and dialkylaminosulfonyl.

R^(b) can be hydrogen, alkyl, haloalkyl, alkenyl, alkynyl, cycloalkyl,cycloalkenyl, aryl, is heteroaryl, or heterocyclyl; wherein each ofalkyl, cycloalkyl, cycloalkenyl, aryl, heteroaryl and heterocycle areoptionally substituted with 1 to 5 substituents independently selectedfrom the group consisting of halo, oxo, —CN, —CHO, —CF₃, —OH, —NO₂,alkyl, —OCF₃, alkoxy, cycloalkoxy, cycloalkenoxy, amino, alkylamino,dialkylamino, acylamino, aminoacyl, alkylsulfonyl, alkylaminosulfonyl,and dialkylaminosulfonyl. In some circumstances, R^(b) and Z⁴ are takento together to form —C(O)O— or ═C(R^(f))O—.

R^(c) can be alkyl, aryl, trifluoromethyl, methylsulfonyl,trifluoromethylsulfonyl, p-tolylsulfonyl, or a group selected such that—OCOR^(c) is a good leaving group;

Each R^(f), independently, is hydrogen, alkyl, haloalkyl, alkenyl,alkynyl, cycloalkyl, cycloalkenyl, aryl, heteroaryl, or heterocyclyl;wherein each of alkyl, cycloalkyl, cycloalkenyl, aryl, heteroaryl andheterocycle are optionally substituted with 1 to 5 substituentsindependently selected from the group consisting of halo, oxo, —CN,—CHO, —CF₃, —OH, —NO₂, alkyl, —OCF₃, alkoxy, cycloalkoxy, cycloalkenoxy,amino, alkylamino, dialkylamino, acylamino, aminoacyl, alkylsulfonyl,alkylaminosulfonyl, and dialkylaminosulfonyl.

Each R^(g), independently, is hydrogen, alkyl, haloalkyl, alkenyl,alkynyl, cycloalkyl, cycloalkenyl, aryl, heteroaryl, or heterocyclyl;wherein each of alkyl, cycloalkyl, cycloalkenyl, aryl, heteroaryl andheterocycle are optionally substituted with 1 to 5 substituentsindependently selected from the group consisting of halo, oxo, —CN,—CHO, —CF₃, —OH, —NO₂, alkyl, —OCF₃, alkoxy, cycloalkoxy, cycloalkenoxy,amino, alkylamino, dialkylamino, acylamino, aminoacyl, alkylsulfonyl,alkylaminosulfonyl, and dialkylaminosulfonyl.

The compound can be in the form of a pharmaceutically acceptable salt orprodrug.

In some embodiments, W is —O—. R^(a) and R^(b), independently, can eachbe H or alkyl. Y can be —OR^(f), or in some circumstances, Y can be —OHor —O—P(O)(OR^(f))OR^(g). X⁶ can be H, halo, alkyl, cycloalkyl, orhaloalkyl.

Simultaneously, A³ can be —C(X³)H—, A⁴ can be —C(X⁴)H—, and A⁵ can be—C(X⁵)H—.

In some embodiments, Cy has the formula:

in which Z¹ is a bond, —[C(R^(d)R^(e))]_(x)—, —CR^(d)═CR^(e)—, —O—,—NR^(f)—; Z² is a bond, —[C(R^(d)R^(e))]_(y)—, —CR^(d)═CR^(e)—, —O—,—NR^(f)—; Z³ is a bond, —[C(R^(d)R^(e))]_(z)—, —CR^(d)═CR^(e)—, —O—,—NR^(f)—; and each of x, y, and z, independently, is 1 to 3.

Each R^(d), independently, can be H, halo, hydroxy, alkyl, haloalkyl,alkenyl, alkoxy, cycloalkyl, —C(O)NR^(f)R^(g), —NR^(f)R^(g),—NR^(f)C(O)R^(g), or —SO₂NR^(f)R^(g).

Each R^(e), independently, can be H, halo, hydroxy, alkyl, haloalkyl,alkenyl, alkoxy, cycloalkyl, —C(O)NR^(f)R^(g), —NR^(f)R^(g),—NR^(f)C(O)R^(g), or —SO₂NR^(f)R^(g).

R^(1a) and R^(1b), independently, can be hydrogen, halo, hydroxy, nitro,cyano, —NR^(f)R^(g), alkyl, haloalkyl, cycloalkyl, cycloalkenyl,cycloalkylalkyl, cycloalkenylalkyl, heterocyclylalkyl, arylalkyl,heteroarylalkyl, alkoxy, cycloalkylalkoxy, cycloalkenylalkoxy,heterocyclylalkoxy, arylalkoxy, heteroarylalkoxy, acyl, cycloalkylacyl,cycloalkenylacyl, heterocyclylacyl, arylacyl, heteroarylacyl, thioalkyl,alkenyl, alkynyl, cycloalkenyl, heterocyclyl, aryl, or heteroaryl. Insome circumstances, R^(1a) and R^(1b), when taken together, can be C₂-0₅alkylene optionally terminated by or inturrepted by 1 or 2 oxygen atoms,or C₂-C₅ alkenylene optionally terminated by or inturrepted by 1 or 2oxygen atoms.

R^(2a) and R^(2b), independently, can be hydrogen, halo, hydroxy, nitro,cyano, —NR^(f)R^(g), alkyl, haloalkyl, cycloalkyl, cycloalkenyl,cycloalkylalkyl, cycloalkenylalkyl, heterocyclylalkyl, arylalkyl,heteroarylalkyl, alkoxy, cycloalkylalkoxy, cycloalkenylalkoxy,heterocyclylalkoxy, arylalkoxy, heteroarylalkoxy, acyl, cycloalkylacyl,cycloalkenylacyl, heterocyclylacyl, arylacyl, heteroarylacyl, thioalkyl,alkenyl, alkynyl, cycloalkenyl, heterocyclyl, aryl, or heteroaryl. Insome circumstances, R^(1a) and R^(2a), when taken together, can be C₁-C₅alkylene optionally terminated by or inturrepted by 1 or 2 oxygen atoms,or C₂-C₅ alkenylene optionally terminated by or inturrepted by 1 or 2oxygen atoms.

R^(1a), R^(1b), R^(2a), and R^(2b) can each, independently, besubstituted with 0-5 substituents selected from halo, hydroxy, nitro,cyano, —NR^(f)R^(g), or —CO₂R^(f).

In some embodiments, R^(1a) and R^(2a) can both be hydrogen. Z¹ can be—CH₂CH₂—. Z² can be —CH₂—. Z³ can be a bond.

R^(1b) can be fluoro, chloro, bromo, iodo, methyl, difluoromethyl,triflurormethyl, ethyl, 1,1-difluoroethyl, propyl, isopropyl, n-butyl,i-butyl, t-butyl, n-pentyl, isopentyl, 1,1-dimethylpropyl, neopentyl,cyclopentyl, n-hexyl, cyclohexyl, methoxy, trifluoromethoxy, ethoxy,n-propoxy, i-propoxy, n-butoxy, i-butoxy, t-butoxy, n-pentyloxy,i-pentyloxy, 1,1-dimethylpropoxy, neopentyloxy, cyclopentyloxy,n-hexyloxy, or cyclohexyloxy.

In another aspect, a compound of formula (II):

in which each of X¹, X², X³, X^(3′), X⁴, X^(4′), X⁵, X^(5′), X⁶, Y, andZ⁴ are defined for formula (I).

R^(1a) and R^(1b), independently, can each be hydrogen, halo, hydroxy,nitro, cyano, —NR^(f)R^(g), alkyl, haloalkyl, cycloalkyl, cycloalkenyl,cycloalkylalkyl, cycloalkenylalkyl, heterocyclylalkyl, arylalkyl,heteroarylalkyl, alkoxy, cycloalkylalkoxy, cycloalkenylalkoxy,heterocyclylalkoxy, arylalkoxy, heteroarylalkoxy, acyl, cycloalkylacyl,cycloalkenylacyl, heterocyclylacyl, arylacyl, heteroarylacyl, thioalkyl,alkenyl, alkynyl, cycloalkenyl, heterocyclyl, aryl, or heteroaryl.

R^(1a) and R^(1b), when taken together, can be C₂-C₅ alkylene optionallyterminated by or inturrepted by 1 or 2 oxygen atoms, or C₂-C₅ alkenyleneoptionally terminated by or inturrepted by 1 or 2 oxygen atoms.

Z¹ can be a bond, —[C(R^(d)R^(e))]_(x)—, or —CR^(d)═CR^(e)—; Z² can be abond, —C[R^(d)R³)]_(y)—, or —CR^(d)═CR^(e)—; and each of x and y,independently, can be 1 to 3.

Each R^(d), independently, can be H, halo, hydroxy, alkyl, alkenyl,alkoxy, or cycloalkyl. Each R^(e), independently, can be H, halo,hydroxy, alkyl, alkenyl, alkoxy, or cycloalkyl. The compound can be inthe form of a pharmaceutically acceptable salt or prodrug.

Y can be —OR^(f), or in some circumstances, Y can be —OH or—O—P(O)(OR^(f))OR^(g). X⁶ can be an electron withdrawing group, or insome circumstances, X⁶ can be H, halo, alkyl, cycloalkyl, or haloalkyl.Z¹ can be —CH₂CH₂—, and Z² can be —CH₂CH₂—. R^(1a) can be hydrogen,halo, hydroxy, alkyl, haloalkyl, cycloalkyl, cycloalkylalkyl, arylalkyl,alkoxy, cycloalkylalkoxy, arylalkoxy, or aryl. Simultaneously, Y can be—OH or —OP(O)(OH)₂; Z⁴ can be H or —OH; X¹, X², X³, X⁴, and X⁵ can eachbe H; X^(3′), X^(4′), and X^(5′) can each be H; and X⁶ can be halo,alkyl, cycloalkyl, or haloalkyl.

Z¹ can be —(CH₂)_(x)— and Z² can be —(CH₂)_(y)—. R^(1a) can be alkyl,haloalkyl, cycloalkyl, aryl, or arylalkoxy. The compound of formula (II)having the formula:

i.e., where R^(1a) and the oxygen atom bound to the cyclohexyl ring arein the trans-orientation with respect to one another.

In another aspect, a compound can have formula (IV):

in which each of X¹, X², X³, X^(3′), X⁴, X^(4′), X⁵, X^(5′), X⁶, Y, andZ⁴ are as defined for formula (I).

In formula (IV), A³ can be —N═, A⁴ can be —C(R⁴)═, A⁵ can be —C(R⁵)═,and A⁶ can be —C(R⁶)═. Or A³ can be —C(R³)═, A⁴ can be —N═, A⁵ can be—C(R⁵)═, and A⁶ can be —C(R⁶)═. Or, A³ can be —C(R³)═, A⁴ can be—C(R⁴)═, A⁵ can be —N═, and A⁶ can be —C(R⁶)═. Or, A³ can be —C(R³)═, A⁴can be —C(R⁴)═, A⁵ can be —C(R⁵)═, and A⁶ can be —N═. Or, A³ can be —N═,A⁴ can be —N═, A⁵ can be —C(R⁵)═, and A⁶ can be —C(R⁶)═.

R^(1a) and R^(1b), independently, can each be hydrogen, halo, hydroxy,nitro, cyano, —NR^(f)R^(g), alkyl, haloalkyl, cycloalkyl, cycloalkenyl,cycloalkylalkyl, cycloalkenylalkyl, heterocyclylalkyl, arylalkyl,heteroarylalkyl, alkoxy, cycloalkylalkoxy, cycloalkenylalkoxy,heterocyclylalkoxy, arylalkoxy, heteroarylalkoxy, acyl, cycloalkylacyl,cycloalkenylacyl, heterocyclylacyl, arylacyl, heteroarylacyl, thioalkyl,alkenyl, alkynyl, cycloalkenyl, heterocyclyl, aryl, or heteroaryl.

R^(1a) and R^(1b), when taken together, can be C₂-C₅ alkylene optionallyterminated by or inturrepted by 1 or 2 oxygen atoms, or C₂-C₅ alkenyleneoptionally terminated by or inturrepted by 1 or 2 oxygen atoms.

Z¹ can be a bond, —[C(R^(d)R^(e))]_(x)—, or —CR^(d)═CR^(e)—; Z² can be abond, —[C(R^(d)R^(e))]_(y)—, or —CR^(d)═CR^(e)—; and each of x and y,independently, can be 1 to 3.

Each R^(d), independently, can be H, halo, hydroxy, alkyl, alkenyl,alkoxy, or cycloalkyl. Each R^(e), independently, can be H, halo,hydroxy, alkyl, alkenyl, alkoxy, or cycloalkyl. The compound can be inthe form of a pharmaceutically acceptable salt or prodrug.

In another aspect, a pharmaceutical composition includes apharmaceutically acceptable carrier and a compound of formula (I) asdefined above.

In another aspect, a method of making a compound of formula (I)comprising contacting a compound of formula (III):

with a compound having the formula: Cy-OH.

In formula (III), each of A¹, A², A³, A⁴, A⁵, and A⁶, are as defined forformula (I). R³ can have the formula:

in which Z⁴ is H or —OR^(f) (where R^(f) is as defined in formula (I));R^(a) is as defined in formula (I); and Pg is an amino protecting group.

In the compound have the formula Cy-OH, Cy can be cycloalkyl,cycloalkenyl, heterocyclyl, aryl, or heteroaryl; wherein Cy isoptionally substituted by 1-6 substituents to selected from the groupconsisting of hydrogen, halo, hydroxy, nitro, cyano, —NR^(f)R^(g),alkyl, haloalkyl, cycloalkyl, cycloalkenyl, cycloalkylalkyl,cycloalkenylalkyl, heterocyclylalkyl, arylalkyl, heteroarylalkyl,alkoxy, haloalkoxy, cycloalkylalkoxy, cycloalkenylalkoxy,heterocyclylalkoxy, aryloxy, arylalkoxy, heteroaryloxy,heteroarylalkoxy, acyl, cycloalkylacyl, cycloalkenylacyl,heterocyclylacyl, arylacyl, heteroarylacyl, thioalkyl, alkenyl, alkynyl,is cycloalkenyl, heterocyclyl, aryl, and heteroaryl. R^(f) and R^(g) areas defined for formula (I).

In another aspect, a method for prevention or treatment of apathological condition or symptom in a mammal, wherein the activity ofsphingosine 1-phosphate receptors is implicated and agonism of suchactivity is desired, includes administering to said mammal an effectiveamount of a compound of formula (I).

The pathological condition can be neuropathic pain. The pathologicalcondition can be an autoimmune disease. The method can includeadministering to said mammal an effective amount of a drug selected fromthe group consisting of: corticosteroids, bronchodilators,antiasthmatics, antiinflammatories, antirheumatics, immunosuppressants,antimetabolites, immunomodulators, antipsoriatics, and antidiabetics.The autoimmune disease is uveitis, type I diabetes, rheumatoidarthritis, inflammatory bowel diseases, lupus, asthma, psoriasis, ormultiple sclerosis.

The prevention or treatment of the pathological condition can includealtering lymphocyte trafficking. Altering lymphocyte trafficking canprovide prolonged allograft survival. The allograft can be fortransplantation.

In another aspect, a method for prevention or treatment of apathological condition or symptom in a mammal, wherein the activity S1Plyase implicated and inhibition of the S1P lyase is desired, includesadministering to said mammal an effective amount of a compound offormula (I).

In another aspect, an assay includes transfecting HEK293 cells with aplasmid encoding sphingosine kinase 2, obtaining a soluble cell lysateincluding sphingosine kinase 2, contacting the soluble cell lysate withATP and a test compound, and determining whether the test compound isphosphorylated.

The details of one or more embodiments are set forth in the accompanyingdescription is below. Other features, objects, and advantages will beapparent from the description and drawings, and from the claims.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a schematic depiction of a synthesis of an intermediate usefulin the preparation of compounds of Formula (I).

FIG. 2 is an illustration of an X-ray crystal structure of anintermediate useful in the preparation of compounds of Formula (I).

FIG. 3 is a schematic depiction of a synthesis of compounds of Formula(I).

FIGS. 4-10 are schematic depictions of syntheses of compounds of Formula(I).

FIG. 11 is a graph depicting the results of various assays on compoundsof formula (I).

DETAILED DESCRIPTION

The following abbreviations are used herein: S1P,sphingosine-1-phosphate; S1P₁₋₅ S1P receptor types; GPCR, G-proteincoupled receptor; SAR, structure-activity relationship; EDG, endothelialcell differentiation gene; EAE, experimental autoimmuneencephalomyelitis; NOD non-obese diabetic; TNFα, tumor necrosis factoralpha; HDL, high density lipoprotein; and RT-PCR, reverse transcriptasepolymerase chain reaction.

The values listed below for radicals, substituents, and ranges, are forillustration only; they do not exclude other defined values or othervalues within defined ranges for the radicals and substituents. Thedisclosed compounds include compounds of formula I having anycombination of the values, specific values, more specific values, andpreferred values described herein.

The term “halogen” or “halo” includes bromo, chloro, fluoro, and iodo.The term “haloalkyl”, refers to an alkyl radical bearing at least onehalogen substituent, non-limiting examples include, but are not limitedto, chloromethyl, fluoroethyl, trichloromethyl, trifluoromethyl and thelike.

The term “C₁-C₂₀ alkyl” refers to a branched or linear alkyl grouphaving from one to twenty carbons. Non-limiting examples include, butare not limited to, methyl, ethyl, n-propyl, iso-propyl, butyl,iso-butyl, sec-butyl, tert-butyl, pentyl, hexyl, heptyl, octyl and thelike.

The term “C₂-C₂₀ alkenyl”, refers to an olefinically unsaturatedbranched or linear group having from two to twenty carbon atoms and atleast one double bond. Typically, C₂-C₂₀ alkenyl groups include, but arenot limited to, 1-propenyl, 2-propenyl, 1,3-butadienyl, 1-butenyl,hexenyl, pentenyl, hexenyl, heptenyl, octenyl and the like.

The term (C₂-C₂₀)alkynyl can be ethynyl, 1-propynyl, 2-propynyl,1-butynyl, 2-butynyl, 3-butynyl, 1-pentynyl, 2-pentynyl, 3-pentynyl,4-pentynyl, 1- hexynyl, 2-hexynyl, 3-hexynyl, 4-hexynyl, or 5-hexynyl,and the like.

The term “(C₁-C₁₀)alkoxy” refers to an alkyl group attached through anoxygen atom. Examples of (C₁-C₁₀)alkoxy can be methoxy, ethoxy, propoxy,isopropoxy, butoxy, iso-butoxy, sec-butoxy, pentoxy, 3-pentoxy, orhexyloxy and the like.

The term “C₃-C₁₂ cycloalkyl” refers to a cyclic alkyl group, such as,for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,cycloheptyl, cyclooctyl and the like. Cyloalkyl groups include bicyclicgroups such as decalinyl, bridged bicyclic groups such as norbornyl andbicyclo[2.2.2]octyl, tricyclic, bridged tricyclic such as adamantyl, andspiro-linked bicyclic or tricyclic groups.

The term “(C₆-C₁₄)aryl” refers to a monocyclic, bicyclic, or tricycliccarbocyclic ring system having one or two aromatic rings including, butnot limited to, phenyl, benzyl, naphthyl, tetrahydronaphthyl, indanyl,indenyl, anthracyl, and the like.

The term “aryl(C₁-C₂₀)alkyl” or “arylalkyl” or “aralkyl” refers to analkyl group substituted with a mono or bicyclic carbocyclic ring systemhaving one or two aromatic rings including, a group such as phenyl,naphthyl, tetrahydronaphthyl, indanyl, indenyl, and the like.Non-limiting examples of arylalkyl include benzyl, phenylethyl, and thelike.

The term “(C₁-C₁₄)heterocyclic group” refers to an optionallysubstituted mono- or bicyclic carbocyclic ring system containing one,two, three, or four heteroatoms (optionally in each ring) wherein theheteroatoms are oxygen, sulfur, and nitrogen.

The term “(C₄-C₁₄)heteroaryl” refers to an optionally substituted mono-or bicyclic cyclic ring system containing one, two, or three heteroatoms(optionally in each ring) wherein the heteroatoms are oxygen, sulfur,and nitrogen. Non-limiting examples of heteroaryl groups include furyl,thienyl, pyridyl, and the like.

The term “phosphate analog” and “phosphonate analog” comprise analogs ofphosphate and phosphonate wherein the phosphorous atom is in the +5oxidation state and one or more of the oxygen atoms is replaced with anon-oxygen moiety, including for example, the phosphate analogsphosphorothioate, phosphorodithioate, phosphoroselenoate,phosphorodiselenoate, phosphoroanilothioate, phosphoranilidate,phosphoramidate, boronophosphates, and the like, including associatedcounterions, e.g., H⁺, NH₄ ⁺, Na⁺, K⁺, and the like if such counterionsare present.

The term “alpha-substituted phosphonate” includes phosphonate(—CH₂PO₃H₂) groups that are substituted on the alpha-carbon such as—CHFPO₃H₂, —CF₂PO₃H₂, —CHOHPO₃H₂, —C═OPO₃H₂) and the like.

The term “pharmaceutically acceptable carrier” includes any of thestandard pharmaceutical carriers, such as a phosphate buffered salinesolution, hydroxypropyl beta-cyclodextrins (HO-propyl betacyclodextrins), water, emulsions such as an oil/water or water/oilemulsion, and various types of wetting agents. The term also encompassesany of the agents approved by a regulatory agency of the U.S. Federalgovernment or listed in the U.S. Pharmacopeia for use in animals,including humans.

The term “pharmaceutically acceptable salt or prodrug” refers to saltswhich retain the biological effectiveness and properties of thedisclosed compounds and which are not biologically or otherwiseundesirable. In many cases, the disclosed compounds are capable offorming acid or base salts by virtue of the presence of amino orcarboxyl groups or groups similar thereto. “Prodrug” refers to acompound that can hydrolyze, oxidize, be phosphorylated, or otherwisereact under biological conditions (in vitro or in vivo) to provide acompound in pharmacologically active form. In the present context, acompound of Formula (I) can be pharmacologically active (e.g., functionas an S1P receptor agonist) when group Y includes, for example, aphosphate group. Suitable prodrugs can therefore include phosphateesters (hydrolyzed to the corresponding phosphate), alcohols(phosphorylated to the corresponding phosphate), esters (hydrolyzed toproduce an alcohol, which is phosphorylated to the correspondingphosphate), oxetanes (e.g., where L¹, Z⁴, Y, and the atoms to which theyare bound form an oxetane ring; the oxetane can be hydrolyzed to producean alcohol, which is phosphorylated to the corresponding phosphate) andother compounds that can be converted to a pharmacologically activeform.

An “effective amount” means an amount sufficient to produce a selectedeffect. For example, an effective amount of an S1P receptor agonist isan amount that decreases the cell signaling activity of the S1Preceptor.

The disclosed compounds can contain one or more asymmetric centers inthe molecule. In accordance with the present disclosure any structurethat does not designate the stereochemistry is to be understood asembracing all the various optical isomers, as well as racemic mixturesthereof.

The disclosed compounds may exist in tautomeric forms and the inventionincludes both mixtures and separate individual tautomers. For example,the following structure:

is understood to represent a mixture of the structures:

as well as

FIGS. 1 and 3 illustrate schematically a synthetic route to certaincompounds of Formula (I), starting from 6-methoxy-1-tetralone. First the2-position is modified by bromination and diethyl methylmalonate.Reduction converts the oxo group to a methylene group. Amination and afurther reduction affords the beta amino alcohol, which is protected asa cyclic carbamate. Then the 6-hydroxy group is unmasked. At this point,the different stereoisomers may be separated (e.g., by chiralchromatography), so that further steps can be carried out usingsubstantially stereopure materials. A Mitsunobu reaction can be used toinstall the Cy group. Finally, the cyclic carbamate protection isreversed to afford the final product.

FIGS. 4 and 5 illustrate schematically a synthetic route to certaincompounds of Formula (I), starting from m-aminophenol or p-aminophenol.Briefly, the starting material is acetylated and subsequently oxidizedto afford 7-acetoxy-2-chloroquinoline-3-carbaldehyde (starting fromm-aminophenol, FIG. 4) or 6-acetoxy-3-chloroquinoline-2-carbaldehyde(starting from p-aminophenol, FIG. 5). The acetoxy group is deprotectedand the chloro group replaced by H. The resulting alcohol can bealkylated at this stage, for example using an alkyl halide. Next aGringnard reaction followed by an oxidation affords a ketone. The ketoneis reacted is with ammonium carbonate, hydrolyzed, and reduced, to givethe final product, a 2-amino-2-(7-alkoxyquinolin-3-yl)-1-propanol as amixture of stereoisomers. If desired, the stereoisomers may beseparated, for example by chiral chromatography.

FIGS. 6-10 illustrate schematically synthetic routes to certaincompounds of Formula (I), e.g., quinolines (FIGS. 6 and 9), quinazolines(FIGS. 7-8) and benzothiazoles (FIG. 10).

An “S1P modulating agent” refers a compound or composition that iscapable of inducing a detectable change in S1P receptor activity in vivoor in vitro (e.g., at least 10% increase or decrease in S1P activity asmeasured by a given assay such as the bioassay described in the examplesand known in the art. “S1P receptor,” refers to all of the S1P receptorsubtypes (for example, the S1P receptors S1P₁, S1P₂, S1P₃, S1P₄, andS1P₅), unless the specific subtype is indicated.

It will be appreciated by those skilled in the art that the disclosedcompounds having chiral centers may exist in and be isolated inoptically active and racemic forms. It is to be understood that thedisclosed compounds encompass any racemic, optically active orstereoisomeric form, or mixtures thereof. It is well known in the arthow to prepare such optically active forms (for example, resolution ofthe racemic form by recrystallization techniques, synthesis fromoptically-active starting materials, by chiral synthesis, orchromatographic separation using a chiral stationary phase) and how todetermine S1P agonist activity using the standard tests describedherein, or using other similar tests which are well known in the art. Inaddition, some compounds may exhibit polymorphism.

In some embodiments, the carbon atom labeled with a * in Formula (I)below can be a stereogenic center.

In such embodiments, there can be a preferred stereochemicalconfiguration. For example, when L² is a bond, L¹ is —CH₂—, Z⁴ is H, andY is —OH, the preferred configuration is the R configuration:

Potential uses of an S1P receptor agonist, and S1P₁ receptor typeselective agonists particularly, include, but are not limited to,altering lymphocyte trafficking as a method of treatment for neuropathicpain, inflammation-induced pain (e.g., where prostaglandins areinvolved) or treatment of autoimmune pathologies such as uveitis, type Idiabetes, rheumatoid arthritis, inflammatory bowel diseases (e.g.,Crohn's disease and ulcerative colitis), multiple sclerosis, lupus,asthma, psoriasis, and in drug-eluting stents. Additional uses caninclude treatment of brain degenerative diseases, heart diseases,cancers, or hepatitis C. See, for example, WO 2005/085295, WO2004/010987, WO 03/097028, and WO 2006/072562, each of which isincorporated by reference in its entirety.

“Treatment” of multiple sclerosis includes treating various forms of thedisease including relapsing-remitting, chronic progressive, and the S1Preceptor agonists can be used alone or in conjunction with other agentsto relieve signs and symptoms of the disease as well asprophylactically.

In addition, the disclosed compounds can be used for altering lymphocytetrafficking as a method for prolonging allograft survival, for examplesolid organ transplants, treatment of graft vs. host disease, bonemarrow transplantation, and the like.

In addition, the disclosed compounds can be used to inhibit autotaxin.Autotaxin, a plasma phosphodiesterase, has been demonstrated to undergoend product inhibition. Autotaxin hydrolyzes several substrates to yieldlysophosphatidic acid and sphingosine 1-phosphate, and has beenimplicated in cancer progression and angiogenesis. Therefore, S1Preceptor agonist pro-drugs of the disclosed compounds can be used toinhibit autotaxin. This activity may be combined with agonism at S1Preceptors or may be independent of such activity.

In addition, disclosed compounds can be useful for inhibition of S1Plyase. S1P lyase is an intracellular enzyme that irreversibly degradesS1P. Inhibition of S1P lyase disrupts lymphocyte trafficking withconcomitant lymphopenia. Accordingly, S1P lyase inhibitors can be usefulin modulating immune system function. Therefore, the disclosed compoundscan be used to inhibit S1P lyase. This inhibition could be in concertwith S1P receptor activity, or be independent of activity at any S1Preceptor.

In addition, disclosed compounds can be useful as antagonists of thecannabinoid CB₁ receptor. CB₁ antagonism is associated with a decreasein body weight and an improvement in blood lipid profiles. The CB₁antagonism could be in concert with S1P receptor activity, or beindependent of activity at any S1P receptor.

In addition, disclosed compounds can be useful for inhibition of groupIVA cytosolic PLA₂ (cPLA₂). cPLA₂ catalyzes the release of eicosanoicacids (e.g., arachidonic acid). The eicosanoic acids are transformed topro-inflammatory eicosanoids such as prostaglandins and leukotrienes.Thus, disclosed compounds may be useful as anti-inflammatory agents.This inhibition could be in concert with S1P receptor activity, or beindependent of activity at any S1P receptor.

In addition, disclosed compounds may be useful for inhibition of themultiple substrate lipid kinase (MuLK). MuLK is highly expressed in manyhuman tumor cells and thus its inhibition might slow the growth orspread of tumors.

Pharmaceutical compositions can include the compounds of formula I. Moreparticularly, such compounds can be formulated as pharmaceuticalcompositions using standard pharmaceutically acceptable carriers,fillers, solubilizing agents and stabilizers known to those skilled inthe art. For example, a pharmaceutical composition including a compoundof formula I, or a salt, analog, derivative, or modification thereof, asdescribed herein, is used to administer the appropriate compound to asubject.

The compounds of formula I are useful for treating a disease or disorderincluding administering to a subject in need thereof of atherapeutically acceptable amount of a compound of formula I, or apharmaceutical composition comprising a therapeutically effective amountof a compound of formula I, and a pharmaceutically-acceptable carrier.

The disclosed compounds and method are directed to sphingosine1-phosphate (S1P) analogs that have activity as receptor receptoragonists or antagonists at one or more S1P receptors, specifically theS1P₁, S1P₄ and S1P₅ receptor types. The disclosed compounds and methodinclude both compounds that have a phosphate moiety as well as compoundswith hydrolysis-resistant phosphate surrogates such as phosphonates,alpha-substituted phosphonates particularly where the alpha substitutionis a halogen and phosphothionates.

The values listed below for radicals, substituents, and ranges, are forillustration only; they do not exclude other defined values or othervalues within defined ranges for the radicals and substituents.

In cases where compounds of formula I are sufficiently basic or acidicto form stable nontoxic acid or base salts, preparation andadministration of the compounds as pharmaceutically acceptable salts maybe appropriate. Examples of pharmaceutically acceptable salts areorganic acid addition salts formed with acids which form a physiologicalacceptable anion, for example, tosylate, methanesulfonate, acetate,citrate, malonate, tartarate, succinate, benzoate, ascorbate,α-ketoglutarate, and α-glycerophosphate. Inorganic salts may also beformed, including hydrochloride, sulfate, nitrate, bicarbonate, andcarbonate salts.

Pharmaceutically acceptable salts may be obtained using standardprocedures well known in the art, for example by reacting a sufficientlybasic compound such as an amine with a suitable acid affording aphysiologically acceptable anion. Alkali metal (for example, sodium,potassium or lithium) or alkaline earth metal (for example calcium)salts of carboxylic acids can also be made.

Pharmaceutically-acceptable base addition salts can be prepared frominorganic and organic bases. Salts from inorganic bases, include but arenot limited to, sodium, potassium, lithium, ammonium, calcium andmagnesium salts. Salts derived from organic bases include, but are notlimited to, salts of primary, secondary and tertiary amines, such asalkyl amines, dialkyl amines, trialkyl amines, substituted alkyl amines,di(substituted alkyl)amines, tri(substituted alkyl)amines, alkenylamines, dialkenyl amines, trialkenyl amines, substituted alkenyl amines,di(substituted alkenyl)amines, tri(substituted alkenyl)amines,cycloalkyl amines, di(cycloalkyl)amines, tri(cycloalkyl)amines,substituted cycloalkyl amines, disubstituted cycloalkyl amine,trisubstituted cycloalkyl amines, cycloalkenyl amines,di(cycloalkenyl)amines, tri(cycloalkenyl)amines, substitutedcycloalkenyl amines, disubstituted cycloalkenyl amine, trisubstitutedcycloalkenyl amines, aryl amines, diaryl amines, triaryl amines,heteroaryl amines, diheteroaryl amines, triheteroaryl amines,heterocyclic amines, diheterocyclic amines, triheterocyclic amines,mixed di- and tri-amines where at least two of the substituents on theamine are different and are alkyl, substituted alkyl, alkenyl,substituted alkenyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl,substituted cycloalkenyl, aryl, heteroaryl, or heterocyclic and thelike. Also included are amines where the two or three substituents,together with the amino nitrogen, form a heterocyclic or heteroarylgroup. Non-limiting examples of amines include, isopropylamine,trimethyl amine, diethyl amine, tri(iso-propyl)amine,tri(n-propyl)amine, ethanolamine, 2-dimethylaminoethanol, tromethamine,lysine, arginine, histidine, caffeine, procaine, hydrabamine, choline,betaine, ethylenediamine, glucosamine, N-alkylglucamines, theobromine,purines, piperazine, piperidine, morpholine, N-ethylpiperidine, and thelike. It should also be understood that other carboxylic acidderivatives would be useful, for example, carboxylic acid amides,including carboxamides, lower alkyl carboxamides, dialkyl carboxamides,and the like.

The compounds of formula I can be formulated as pharmaceuticalcompositions and administered to a mammalian host, such as a humanpatient in a variety of forms adapted to the chosen route ofadministration, e.g., orally or parenterally, as eyedrops, byintravenous, intramuscular, topical or subcutaneous routes.

Thus, the present compounds may be systemically administered, e.g.,orally, in combination with a pharmaceutically acceptable vehicle suchas an inert diluent or an assimilable edible carrier. They may beenclosed in hard or soft shell gelatin capsules, may be compressed intotablets, or may be incorporated directly with the food of the patient'sdiet. For oral therapeutic administration, the active compound may becombined with one or more excipients and used in the form of ingestibletablets, buccal tablets, troches, capsules, elixirs, suspensions,syrups, wafers, and the like. Such compositions and preparations shouldcontain at least about 0.1% of active compound. The percentage of thecompositions and preparations may, of course, be varied and mayconveniently be between about 2 to about 60% of the weight of a givenunit dosage form. The amount of active compound in such therapeuticallyuseful compositions is such that an effective dosage level will beobtained.

The tablets, troches, pills, capsules, and the like may also contain thefollowing: binders such as gum tragacanth, acacia, corn starch orgelatin; excipients such as dicalcium phosphate; a disintegrating agentsuch as corn starch, potato starch, alginic acid and the like; alubricant such as magnesium stearate; and a sweetening agent such assucrose, fructose, lactose or aspartame or a flavoring agent such aspeppermint, oil of wintergreen, or cherry flavoring may be added. Whenthe unit dosage form is a capsule, it may contain, in addition tomaterials of the above type, a liquid carrier, such as a vegetable oilor a polyethylene glycol. Various other materials may be present ascoatings or to otherwise modify the physical form of the solid unitdosage form. For instance, tablets, pills, or capsules may be coatedwith gelatin, wax, shellac or sugar and the like. A syrup or elixir maycontain the active compound, sucrose or fructose as a sweetening agent,methyl and propylparabens as preservatives, a dye and flavoring such ascherry or orange flavor. Of course, any material used in preparing anyunit dosage form should be pharmaceutically acceptable and substantiallynon-toxic in the amounts employed. In addition, the active compound maybe incorporated into sustained-release preparations and devices.

The active compound may also be administered intravenously orintraperitoneally by infusion or injection. Solutions of the activecompound or its salts can be prepared in water, optionally mixed with anontoxic surfactant. Dispersions can also be prepared in glycerol,liquid polyethylene glycols, triacetin, and mixtures thereof and inoils. Under ordinary conditions of storage and use, these preparationscontain a preservative to prevent the growth of microorganisms.

Exemplary pharmaceutical dosage forms for injection or infusion caninclude sterile aqueous solutions or dispersions or sterile powderscomprising the active ingredient which are adapted for theextemporaneous preparation of sterile injectable or infusible solutionsor dispersions, optionally encapsulated in liposomes. In all cases, theultimate dosage form should be sterile, fluid and stable under theconditions of manufacture and storage. The liquid carrier or vehicle canbe a solvent or liquid dispersion medium comprising, for example, water,ethanol, a polyol (for example, glycerol, propylene glycol, liquidpolyethylene glycols, and the like), vegetable oils, nontoxic glycerylesters, and mixtures thereof. The proper fluidity can be maintained, forexample, by the formation of liposomes, by the maintenance of therequired particle size in the case of dispersions or by the use ofsurfactants. The prevention of the action of microorganisms can bebrought about by various antibacterial and antifungal agents, forexample, parabens, chlorobutanol, phenol, sorbic acid, thimerosal, andthe like. In many cases, it will be preferable to include isotonicagents, for example, sugars, buffers or sodium chloride. Prolongedabsorption of the injectable compositions can be brought about by theuse in the compositions of agents delaying absorption, for example,aluminum monostearate and gelatin.

Sterile injectable solutions are prepared by incorporating the activecompound in the required amount in the appropriate solvent with variousof the other ingredients enumerated above, as required, followed byfilter sterilization. In the case of sterile powders for the preparationof sterile injectable solutions, the preferred methods of preparationare vacuum drying and the freeze drying techniques, which yield a powderof the active ingredient plus any additional desired ingredient presentin the previously sterile-filtered solutions.

For topical administration, the present compounds may be applied in pureform, e.g., when they are liquids. However, it will generally bedesirable to administer them to the skin as compositions orformulations, in combination with a dermatologically acceptable carrier,which may be a solid or a liquid.

Exemplary solid carriers include finely divided solids such as talc,clay, microcrystalline cellulose, silica, alumina and the like. Usefulliquid carriers include water, alcohols or glycols orwater-alcohol/glycol blends, in which the present compounds can bedissolved or dispersed at effective levels, optionally with the aid ofnon-toxic surfactants. Adjuvants such as fragrances and additionalantimicrobial agents can be added to optimize the properties for a givenuse. The resultant liquid compositions can be applied from absorbentpads, used to impregnate bandages and other dressings, or sprayed ontothe affected area using pump-type or aerosol sprayers.

Thickeners such as synthetic polymers, fatty acids, fatty acid salts andesters, fatty alcohols, modified celluloses or modified mineralmaterials can also be employed with liquid carriers to form spreadablepastes, gels, ointments, soaps, and the like, for application directlyto the skin of the user.

Examples of useful dermatological compositions which can be used todeliver the compounds of formula I to the skin are known to the art; forexample, see Jacquet et al. (U.S. Pat. No. 4,608,392), Geria (U.S. Pat.No. 4,992,478), Smith et al. (U.S. Pat. No. 4,559,157) and Wortzman(U.S. Pat. No. 4,820,508), each of which is incorporated by reference inits entirety.

Useful dosages of the compounds of formula I can be determined bycomparing their in vitro activity, and in vivo activity in animalmodels. Methods for the extrapolation of effective dosages in mice, andother animals, to humans are known to the art; for example, see U.S.Pat. No. 4,938,949, which is incorporated by reference in its entirety.

Generally, the concentration of the compound(s) of formula I in a liquidcomposition, such as a lotion, will be from about 0.1 to about 25 weightpercent, preferably from about 0.5-10 weight percent. The concentrationin a semi-solid or solid composition such as a gel or a powder will beabout 0.1-5 wt-%, preferably about 0.5-2.5 weight percent based on thetotal weight of the composition.

The amount of the compound, or an active salt or derivative thereof,required for use in treatment will vary not only with the particularsalt selected but also with the route of administration, the nature ofthe condition being treated and the age and condition of the patient andwill be ultimately at the discretion of the attendant physician orclinician. In general, however, a dose will be in the range of fromabout 0.1 to about 10 mg/kg of body weight per day.

The compound is conveniently administered in unit dosage form; forexample, containing 0.01 to 10 mg, or 0.05 to 1 mg, of active ingredientper unit dosage form. In some embodiments, a dose of 5 mg/kg or less issuitable.

Ideally, the active ingredient should be administered to achieve peakplasma concentrations of the active compound of from about 1 nm to about50 μM, preferably, about 10 nM to 5 μM, most preferably, about 10 nM toabout 1 μM. This may be achieved, for example, by the intravenousinjection of a 0.05 to 5% solution of the active ingredient, optionallyin saline, or orally administered as a bolus containing about 0.01 to 10μg of the active ingredient. Desirable blood levels may be maintained bycontinuous infusion to provide about 0.01-5.0 mg/kg/hr or byintermittent infusions containing about 0.4-15 mg/kg of the activeingredient(s).

The desired dose may conveniently be presented in a single dose or asdivided doses administered at appropriate intervals, for example, astwo, three, four, or more sub-doses per day, or more infrequently, suchas one to five times a week, or one to five times a month. The sub-doseitself may be further divided, e.g., into a number of discrete looselyspaced administrations; such as multiple inhalations from an insufflatoror by application of a plurality of drops into the eye.

The disclosed method includes a kit comprising a compound of formula Iand instructional material which describes administering the compound ora composition comprising the compound to a cell or a subject. Thisshould be construed to include other embodiments of kits that are knownto those skilled in the art, such as a kit comprising a (preferablysterile) solvent for dissolving or suspending the compound orcomposition prior to administering the compound or composition to a cellor a subject. Preferably, the subject is a human.

In accordance with the disclosed compounds and methods, as describedabove or as discussed in the Examples below, there can be employedconventional chemical, cellular, histochemical, biochemical, molecularbiology, microbiology, and in vivo techniques which are known to thoseof skill in the art. Such techniques are explained fully in theliterature.

The following working examples are provided for the purpose ofillustration only, and are not to be construed as limiting in any waythe remainder of the disclosure. Therefore, the examples should beconstrued to encompass any and all variations which become evident as aresult of the teaching provided herein.

Examples

General: Reactions were run under an inert atmosphere. The usual work-upprocedure was to add organic solvent, usually ethyl acetate, wash withwater or brine, dried, (usually over anhydrous magnesium sulfate) andsolvent removed under reduced pressure. If necessary, the mixture waspurified by column chromatography.

Example 1 2-bromo-6-methoxy-3,4-dihydronaphthalen-1(2H)-one

A 10 L flask was fitted with two cooling condensers and a dry pipe. 6.75L of 1,2-dichloroethane and 750 grams of 6-methoxy-1-tetralone wereadded and stirred to get a brown transparent liquid. The temperature wasraised to 75° C., then 471 g of CuBr₂ was added over a period of 30 min.The mixture was heated to reflux for 2 hrs and a further 337.5 g ofCuBr₂added. After reflux overnight, the color had changed toyellowish-green-gray. HPLC revealed that 10% starting material stillremained; a further 100 g of CuBr₂ was added and stirred for 3 hours tocomplete the reaction.

The reaction was cooled to room temperature, filtered, and the filtercake was washed with 1,2-dichloroethane (750 mL×2), and the motherliquid was washed with NaHCO₃ (1500 mL×4), then washed with brine (750mL×2). The organic layer was dried with Na₂SO₄ and de-colored withactive carbon. The liquid was maintained at a temperature lower than 90°C., concentrated under vacuum, and cooled to 50° C. Methanol was added,the solution decolorized with charcoal, filtered, and washed with 300 mLmethanol. The liquids were combined, cooled to 0-5° C. for 30 min,filtered, then washed with 150 mL methanol. The cake was dried to obtain940 g of 2-bromo-6-methoxy-3,4-dihydronaphthalen-1(2H)-one, M=255.2,purity 98.48%.

Example 2 Diethyl2-methyl-2-(6-methoxy-1-oxo-1,2,3,4-tetrahydronaphthalen-2-yl)malonate

To 6 L of ice cold DMF was added 183 grams of NaH; the mixture stirredand a solution of 811.2 grams diethyl methylmalonate in 1.5 L DMF wasadded dropwise. The temp was kept at 0-5° C. for 2 hrs, and then cooledbelow −3° C., and 650 gramsbromo-6-methoxy-3,4-dihydronaphthalen-1(2H)-one was added. The reactionwas followed by HPLC until the starting material disappeared. Themixture was poured onto ice water, adjusted to pH 4-5 with HCl, andextracted with ethyl acetate. The organic layer was washed with brine,dried, decolorized and concentrated. To the residue was added ether, themixture cooled, filtered, and the residue washed with petroleum ether toget 745 g of diethyl2-methyl-2-(6-methoxy-1-oxo-1,2,3,4-tetrahydronaphthalen-2-yl)malonate,M=348.2, purity 96.57%.

Example 32-acetamido-3-ethoxy-2-(6-methoxy-1,2,3,4-tetrahydronaphthalen-2-yl)-3-oxopropanoicacid

To 218 g KOH in 2.8 L absolute ethanol was added 560 grams diethyl2-methyl-2-(6-methoxy-1-oxo-1,2,3,4-tetrahydronaphthalen-2-yl)malonate.The mixture was stirred for 15 hrs. When HPLC showed <2% of startingmaterial remained, the solution was poured into ice water, and the flaskwas washed with 560 mL cold water. The pH of the solution was adjustedto 2.0-3.0 with 6N HCl. The solution was extracted with dichloromethanethree times (1680 mL, 1120 mL, 560 mL), and the organic layers werecombined and washed with brine. The temperature was kept <70° C. duringconcentration. The residue was dissolved in dichloromethane, and REDUCER(700 mL dichloromethane, 1.3 L triethylsilane, 179 mL trifluoroaceticacid, 1 L boron trifluoride etherate, and a further 530 mLdichloromethane) was added dropwise and followed by HPLC until complete.Product was 261 g of2-methyl-3-ethoxy-2-(6-methoxy-1,2,3,4-tetrahydronaphthalen-2-yl)-3-oxopropanoicacid, M=320.2, purity 94%.

Example 4 ethyl2-amino-2-(6-methoxy-1,2,3,4-tetrahydronaphthalen-2-yl)propanoate

To 225 g of 2-methyl-3-ethoxy-2-(6-methoxy-1,2,3,4-tetrahydronaphthalen-2-yl)-3-oxopropanoic acid in 2.7 L toluene,315 mL diphenyl phosphoryl azide (DPA) in 207 mL of triethylamine wasadded. The mixture was heated to reflux (100-110° C.) for 2 hrs, andcooled to 0° C. A fresh trimethylsilonate sodium salt solution (made byadding 240 g NaOH to 487 g hexamethyl disiloxane and 3L DME, reflux for20 hrs, filtered, washed with DME 600 mL×2, and concentrated, followedby addition of toluene 2 L×2, and concentrated. The mixture was cooledto 30° C., and 3200 mL THF added and stirred for 30 min to obtain thesodium trimethylsilanolate THF solution) was added dropwise thenstirring was stopped and kept overnight. The solution was added to 10%citric acid, pH 3-4, the water layer was extracted with methyltert-butyl ether three times, organic layers combined, washed withaqueous NaHCO₃, with brine, dried (temperature lower than 90° C.),cooled to room temperature. A mixture of 1.2 L HCl and 3.6 L water wasadded, stirred for 30 min, washed with methyl tert-butyl ether, adjustedthe pH of water layer to 8-9, extracted with methyl tert-butyl ether.The organic layer was dried with Na₂SO₄, decolorized and concentrated toget an oil.

Example 52-amino-2-(6-methoxy-1,2,3,4-tetrahydronaphthalen-2-yl)propan-1-ol

To a 2 L flask cooled by ice water was added 60 g LiAlH₄, and 1 L ofTHF. When the mixture was cooled to below 10° C., a mixture of 118.8grams of ethyl2-amino-2-(6-methoxy-1,2,3,4-tetrahydronaphthalen-2-yl)propanoate and350 mL THF was added dropwise. No starting material remained by HPLC.The temperature was kept below 30° C. while 100 mL ethyl acetate wasadded. To the concentrated mixture was added 400 mL ethyl acetate,followed by 25 mL ice water. The mixture was filtered, and the filtercake with 300 mL ethyl acetate. The combined organic layers washed withbrine 100 mL×2, dried with Na₂SO₄, decolorized with carbon andconcentrated. To the oil, was added ether, the mixture stirred forovernight, filtered and dried to afford 50 g of2-amino-2-(6-methoxy-1,2,3,4-tetrahydronaphthalen-2-yl)propan-1-ol.

Example 64-(6-methoxy-1,2,3,4-tetrahydronaphthalen-2-yl)-4-methyloxazolidin-2-one

To 25 grams of2-amino-2-(6-methoxy-1,2,3,4-tetrahydronaphthalen-2-yl)propan-1-ol in575 mL of dichloromethane was added 45 g of triethylamine. The mixturewas cooled to 0° C., and kept below 5° C. while 21 g of triphosgene indichloromethane was added dropwise. The mixture was stirred at 20-25° C.for 1.5 hrs. The reaction was followed by HPLC until the startingmaterial disappeared. The mixture was concentrated under reducedpressure, 375 mL ethyl acetate was added to the residue. It was washedwith water (50 mL×2), dried, decolorized with carbon, and concentratedand dried to afford 10 g of4-(6-methoxy-1,2,3,4-tetrahydronaphthalen-2-yl)-4-methyloxazolidin-2-one,M=261, purity 97%.

Example 74-(6-hydroxy-1,2,3,4-tetrahydronaphthalen-2-yl)-4-methyloxazolidin-2-one

A stirred solution of 100 grams of4-(6-methoxy-1,2,3,4-tetrahydronaphthalen-2-yl)-4-methyloxazolidin-2-onein 6 L dichloromethane was cooled to −80° C. under nitrogen. A solutionof 177 g BBr₃ in dichloromethane was added dropwise; during addition thereaction temperature was kept below −75° C. The reaction was stirred at20° C. until the starting material was less than 2% by HPLC. Ice waterwas added to crystallize the solid, which was was filtered, washed withwater, and the cake was slurried in methanol and dried to afford 80 g of4-(6-hydroxy-1,2,3,4-tetrahydronaphthalen-2-yl)-4-methyloxazolidin-2-one,M=247.29, purity >98%.

Example 8(4S)-4-((2S)-5-Bromo-6-hydroxy-1,2,3,4-tetrahydronaphthalen-2-yl)-4-methyloxazolidin-2-one

Stereoisomers of4-(6-hydroxy-1,2,3,4-tetrahydronaphthalen-2-yl)-4-methyloxazolidin-2-one(7 g) were separated using superfluid chromatography with two stages.First, using Chiralpak AS-H (2×25 cm) 07-8656 (30% methanol/CO₂, 100bar, 70 mL/min, 220 nm. inj vol.: 1.5 mL, 20 mg/mL 1:1 ethanol:DCM)yielded 1.45 g of diastereomer-1 (R_(f)=5.32 min, chemical purity>99%,de>99%), 1.52 g of diastereomer-2 (R_(f)=6.26 min, chemical purity>99%,de>99%), and the mixture of isomers 3 and 4. Next, isomers 3 and 4 wereseparated using Chiralpak IC (3×15 cm) 806271 (30% isopropanol/CO₂, 100bar, 75 mL/min, 220 nm., inj vol.: 1.25 mL, 35 mg/mL methanol) yielded1.28 g of diastereomer-3 (R_(f)=7.97 min, chemical purity>99%, de>99%)and 1.13 g of diastereomer-4(R_(f)=8.96 min, chemical purity>99%,de>99%). ¹H NMR showed that isomer 1 and 4 were an enantiomeric pair,and isomer 2 and 3 were another enantiomeric pair.

To a mixture of4-(6-hydroxy-1,2,3,4-tetrahydronaphthalen-2-yl)-4-methyloxazolidin-2-one(100 mg, 0.0004 mol) (pure isomer 2) in N,N-Dimethylformamide (0.5 mL,0.006 mol) was added N-Bromosuccinimide (72.0 mg, 0.000404 mol) inN,N-Dimethylformamide (0.5 mL, 0.006 mol) and was stirred at for 6 hrs.Water was added, and the mixture was extracted with dichloromethane. Thesolvent was removed and residue was purified with Isco (12 g silica gel,0-40% MeOH:DCM) to give a white precipitate (125 mg, 95% yield). ¹H NMRshowed a single isomer. The solid (10.1 mg) was dissolved in MeOH (3 mL)and the solvent allowed to evaporate slowly to give white needles, whichwas washed with MeOH and collected (5.0 mg). LCMS 1.23 min 328.21([M+2], 100%). X-ray shows (S,S) configuration, see FIG. 2.

To obtain the X-ray crystal structure, one of the prisms was cut to 0.03mm×0.10 mm×0.10 mm in size, mounted on a nylon loop with Paratone-N oil,and transferred to a Bruker SMART APEX II diffractometer equipped withan Oxford Cryosystems 700 Series Cryostream Cooler and Mo Kα radiation(λ=0.71073 Å). A total of 1823 frames were collected at 193 (2) K toθ_(max)=27.50° with an ω oscillation range of 0.5°/frame, and anexposure time of 40 s/frame using the APEX2 suite of software. (BrukerAXS, 2006a) Unit cell refinement on all observed reflections, and datareduction with corrections for Lp and decay were performed using SAINT.(Bruker AXS, 2006b) Scaling and a numerical absorption correction weredone using SADABS. (Bruker AXS, 2004) The minimum and maximumtransmission factors were 0.7429 and 0.9112, respectively. A total of21361 reflections were collected, 2977 were unique (R_(int)=0.0499), and2708 had I>2σ(I). Systematic absences were consistent with the compoundhaving crystallized in the orthorhombic space group P2₁2₁2₁ (No. 19).The observed mean |E²−1| value was 0.687 (versus the expectation valuesof 0.968 and 0.736 for centric and noncentric data, respectively).

The structure was solved by direct methods and refined by full-matrixleast-squares on F² using SHELXTL. (Bruker AXS, 2001) The asymmetricunit was found to contain one molecule of(4S)-4-((2S)-5-bromo-6-hydroxy-1,2,3,4-tetrahydronaphthalen-2-yl)-4-methyloxazolidin-2-one.All of the nonhydrogen atoms were refined with anisotropic displacementcoefficients. The hydrogen atoms were assigned isotropic displacementcoefficients U(H)=1.2U(C), 1.5U(C_(methyl)), 1.5 U(N) or 1.5U(O), andtheir coordinates were allowed to ride on their respective carbons,nitrogen or oxygen. The refinement converged to R(F)=0.0340,wR(F²)=0.0908, and S=1.084 for 2708 reflections with I>2σ(I), andR(F)=0.0392, wR(F²)=0.0931, and S=1.084 for 2977 unique reflections and173 parameters. The maximum in the final cycle of least-squares was0.001, and the residual peaks on the final difference-Fourier map rangedfrom −0.286 to 0.659 eÅ⁻³. Scattering factors were taken from theInternational Tables for Crystallography, Volume C. (Maslen et al.,1992, and Creagh & McAuley, 1992).

The Flack absolute structure parameter refined to x=−0.001 (12) (versusthe expectation values of 0 (within 3 esd's) for the correct and +1 forthe inverted absolute structure) indicating that the coordinates werefor the correct hand (i.e., (4S)(2S)). (Flack, 1983) For comparison, theincorrect (4R)(2R) enantiomer gave R(F)=0.0628, wR(F²)=0.1627, andS=1.034 for 2708 reflections with I>2σ(I), and R(F)=0.0681,wR(F²)=0.1664, and S=1.034 for 2977 unique reflections and 173parameters. The Flack parameter for that incorrect enantiomer was x=1.01(2).

Example 9(S)-2-amino-2-((R)-6-(trans-4-tert-butylcyclohexyloxy)-1,2,3,4-tetrahydronaphthalen-2-yl)propan-1-ol

The mixture of cis-4-tert-butylcyclohexanol (75.8 mg, 0.000485 mol),(S)-4-((R)-6-hydroxy-1,2,3,4-tetrahydronaphthalen-2-yl)-4-methyloxazolidin-2-one(100 mg, 0.0004 mol) (isomer 1) and triphenylphosphine (127 mg, 0.000485mol) in tetrahydrofuran (4 mL, 0.05 mol) was heated to reflux, anddiisopropyl azodicarboxylate (0.0955 mL, 0.000485 mol) was addeddropwise and was stirred and refluxed for overnight. TLC and LCMSmonitoring, 2.29 (386.43, M+1, 60%) showed the reaction to beincomplete. The mixture was taken up into DCM and subjected tochromatography purification with EtOAc/hexane (10:90 to 80:20) to giveproduct (109.3 mg, 70% yield).

The mixture of(S)-4-((R)-6-(trans-4-tert-butylcyclohexyloxy)-1,2,3,4-tetrahydronaphthalen-2-yl)-4-methyloxazolidin-2-one(109.3 mg, 0.0002835 mol) (isomer 1) and lithium hydroxide (74.7 mg,0.00312 mol) in ethanol (1.8 mL, 0.031 mol) and water (0.60 mL, 0.033mol) was heated to reflux for overnight. LCMS showed starting materialwas consumed and peak R_(f)=1.66 min 360.39 ([M+1], 100%) left. Thesolvent was removed under vacuum and the residue was partitioned betweenwater and dichloromethane. The aqueous layer was extensively extractedwith dichloromethane. And the combined organic phase was dried overNa₂SO₄. The concentrated residue was taken up into dichloromethane andsubjected to chromatography purification with dichloromethane/MeOH(10:90 to 80:20) to give the product (17.3 mg, 17% yield). LCMS360.39([M+1], 100%). NMR (400 MHz, CD3OD) δ ppm 0.89 (s, 9H), 1.07 (s, 3H),1.09-1.54 (m, 6H), 1.79-1.87 (m, 3H), 2.04 (m, 1H), 2.15 (m, 2H), 2.53(dd, J=15.5, 12.3 Hz, 1H), 2.72-2.86 (m, 3H), 3.46 (d, J=11.0 Hz, 1H),3.55 (d, J=11.0 Hz, 1H), 4.07 (m, 1H), 6.59 (d, J=2.4 Hz, 1H), 6.63 (dd,J=8.3, 2.5 Hz, 1H), 6.94 (d, J=8.3 Hz, 1H).

Example 10(S)-2-amino-2-((S)-6-(trans-4-tert-butylcyclohexyloxy)-1,2,3,4-tetrahydronaphthalen-2-yl)propan-1-ol

The mixture of cis-4-tert-Butylcyclohexanol (75.8 mg, 0.000485 mol),(S)-4-((S)-6-hydroxy-1,2,3,4-tetrahydronaphthalen-2-yl)-4-methyloxazolidin-2-one(100 mg, 0.0004 mol) (isomer 2) and triphenylphosphine (127 mg, 0.000485mol) in tetrahydrofuran (4 mL, 0.05 mol) was heated to reflux, anddiisopropyl azodicarboxylate (0.0955 mL, 0.000485 mol) was addeddropwise and was stirred and refluxed for overnight. TLC and LCMSmonitoring, 2.28 (386.46, M+1, 40%) showed the reaction to beincomplete. The mixture was taken up into dichloromethane and subjectedto chromatography purification with EtOAc/hexane (10:90 to 80:20) togive product (46.6mg, 30% yield).

The mixture of(S)-4-((S)-6-(trans-4-tert-butylcyclohexyloxy)-1,2,3,4-tetrahydronaphthalen-2-yl)-4-methyloxazolidin-2-one(46.6 mg, 0.000121 mol) (isomer 2) and lithium hydroxide (31.8 mg,0.00133 mol) in ethanol (0.77 mL, 0.013 mol) and water (0.26 mL, 0.014mol) was heated to reflux for overnight. LCMS showed starting materialwas consumed and peak R_(f)=1.66 min left. The solvent was removed undervacuum and the residue was partitioned between water anddichloromethane. The aqueous was extensively extracted withdichloromethane. And the combined organic phase was dried over Na₂SO₄.The concentrated residue was taken up into dichloromethane and subjectedto chromatography purification with dichloromethane/MeOH (10:90 to80:20) to give the product (29.0 mg, 66% yield). LCMS360.42 ([M+1],100%). ¹H NMR (400 MHz, CD3OD) δ ppm 0.89 (s, 9H), 1.06 (s, 3H),1.04-1.40 (m, 6H), 1.77-1.88 (m, 3H), 1.97 (m, 1H), 2.15 (m, 2H), 2.56(dd, J=15.8, 12.1 Hz, 1H), 2.71-2.84 (m, 3H), 3.47 (d, J=10.9 Hz, 1H),3.51 (d, J=10.9 Hz, 1H), 4.07 (m, 1H), 6.58 (d, J=2.5 Hz, 1H), 6.63 (dd,J=8.2, 2.5 Hz, 1H), 6.96 (d, J=8.2 Hz, 1H).

Example 11(R)-2-amino-2-((R)-6-(trans-4-tert-butylcyclohexyloxy)-1,2,3,4-tetrahydronaphthalen-2-yl)propan-1-ol

The mixture of cis-4-tert-butylcyclohexanol (75.8 mg, 0.000485 mol),(R)-4-((R)-6-hydroxy-1,2,3,4-tetrahydronaphthalen-2-yl)-4-methyloxazolidin-2-one(100 mg, 0.0004 mol) (isomer 3) and triphenylphosphine (127 mg, 0.000485mol) in tetrahydrofuran (4 mL, 0.05 mol) was heated to reflux, anddiisopropyl azodicarboxylate (0.0955 mL, 0.000485 mol) was addeddropwise and was stirred and refluxed for overnight. TLC and LCMSmonitoring, 2.28 (386.29, M+1, 40%) showed the reaction to beincomplete. The mixture was taken up into dichloromethane and subjectedto chromatography purification with EtOAc/hexane (10:90 to 80:20) togive product (44.3mg, 30%).

The mixture of(R)-4-((R)-6-(trans-4-tert-butylcyclohexyloxy)-1,2,3,4-tetrahydronaphthalen-2-yl)-4-methyloxazolidin-2-one(46.6 mg, 0.000121 mol) (from isomer 3) and lithium hydroxide (31.8 mg,0.00133 mol) in ethanol (0.77 mL, 0.013 mol) and water (0.26 mL, 0.014mol) was heated to reflux for overnight. LCMS showed SM was consumed andpeak R_(f)=1.65 left. The solvent was removed under vacuum and theresidue was partitioned between water and dichloromethane. The aqueouswas extensively extracted with dichloromethane and the combined organicphase was dried over Na₂SO₄. The concentrated residue was taken up intodichloromethane and subjected to chromatography purification withdichloromethane/MeOH (10:90 to 80:20) to give the product (21.2 mg, 49%yield). LCMS 360.42 ([M+1], 100%). ¹H NMR (400 MHz, CD₃OD) δ ppm 0.89(s, 9H), 1.06 (s, 3H), 1.04-1.40 (m, 6H), 1.77-1.88 (m, 3H), 1.97 (m,1H), 2.15 (m, 2H), 2.56 (dd, J=15.8, 12.1 Hz, 1H), 2.71-2.84 (m, 3H),3.47 (d, J=10.9 Hz, 1H), 3.51 (d, J=10.9 Hz, 1H), 4.07 (m, 1H), 6.58 (d,J=2.5 Hz, 1H), 6.63 (dd, J=8.2, 2.5 Hz, 1H), 6.96 (d, J=8.2 Hz, 1H).

Example 12(R)-2-amino-2-((S)-6-(trans-4-tert-butylcyclohexyloxy)-1,2,3,4-tetrahydronaphthalen-2-yl)propan-1-ol

The mixture of cis-4-tert-butylcyclohexanol (75.8 mg, 0.000485 mol),(R)-4-((S)-6-hydroxy-1,2,3,4-tetrahydronaphthalen-2-yl)-4-methyloxazolidin-2-one(100 mg, 0.0004 mol) (isomer 4) and triphenylphosphine (127 mg, 0.000485mol) in tetrahydrofuran (4 mL, 0.05 mol) was heated to reflux, anddiisopropyl azodicarboxylate (0.0955 mL, 0.000485 mol) was addeddropwise and was stirred and refluxed for overnight. TLC and LCMSmonitoring, 2.28 (386.39, M+1, 50%) showed the reaction to beincomplete. The mixture was taken up into dichloromethane and subjectedto chromatography purification with EtOAc/hexane (10:90 to 80:20) togive product (71.7mg, 40% yield).

The mixture of(R)-4-((S)-6-(trans-4-tert-butylcyclohexyloxy)-1,2,3,4-tetrahydronaphthalen-2-yl)-4-methyloxazolidin-2-one(46.6 mg, 0.000121 mol) (from isomer 4) and lithium hydroxide (31.8 mg,0.00133 mol) in ethanol (0.77 mL, 0.013 mol) and water (0.26 mL, 0.014mol) was heated to reflux for overnight. LCMS showed starting materialwas consumed and peak R_(f)=1.66 min left. The solvent was removed undervacuum and the residue was partitioned between water anddichloromethane. The aqueous layer was extensively extracted withdichloromethane and the combined organic phase was dried over Na₂SO₄.The concentrated residue was taken up into dichloromethane and subjectedto chromatography purification with dichloromethane/MeOH (10:90 to80:20) to give the product. (15.7 mg, 36% yield). LCMS 360.43 ([M+1],100%). NMR (400 MHz, CD3OD) δ ppm 0.89 (s, 9H), 1.07 (s, 3H), 1.09-1.54(m, 6H), 1.79-1.87 (m, 3H), 2.04 (m, 1H), 2.15 (m, 2H), 2.53 (dd,J=15.5, 12.3 Hz, 1H), 2.72-2.86 (m, 3H), 3.46 (d, J=11.0 Hz, 1H), 3.55(d, J=11.0 Hz, 1H), 4.07 (m, 1H), 6.59 (d, J=2.4 Hz, 1H), 6.63 (dd,J=8.3, 2.5 Hz, 1H), 6.94 (d, J=8.3 Hz, 1H).

Example 13 2-amino-2-(6-heptyloxyquinolin-3-yl)-1-propanol

3-aminophenol (19 g, 17 mol) was dissolved in Ac₂O (162 g, 1.59 mol, 9.5eq.), and pyridine (4.9 g, 0.062 mol, 0.36 eq.) was added. Then thereaction mixture was stirred at 80° C. over 2 h. Ice water (50 mL) wasadded to the mixture, and added saturated NaHCO₃ solution to the mixtureuntil pH=7, then extracted (ethyl acetate), washed (brine), dried(Na₂SO₄), concentrated to give 3-acetamidophenyl acetate as gray solid(30 g, yield: 93%). ESI-MS: 194 (M+H)⁺.

A three-neck flask was charged with DMF (25 mL, 0.325 mol, 3 eq.), thenPOCl₃ (70 mL, 0.758 mol, 7 eq.) was added to the DMF at 0° C. Thesolution was stirred at 0° C. for 30 min. Then 3-acetamidophenyl acetate(20.8 g, 0.108 mol) was added to the mixture at 0° C.

After 30 min the mixture heated to 65° C. and stirred for 16 h. Then,the reaction mixture was added ice water (300 mL) and neutralized withsaturated NaHCO₃ to pH=6, extracted (ethyl acetate), washed (brine),dried (Na₂SO₄), filtered and evaporated to dryness to give the crudeproduct, which was purified by silica gel column chromatography (ethylacetate:pet. ether, 3:1) to give 2-chloro-3-formylquinolin-7-yl acetateas gray solid (2.67 g, yield 10%). ESI-MS: 250 (M+H)⁺.

To a solution of 2-chloro-3-formylquinolin-7-yl acetate (2.9 g, 14 mmol)and Pd(PPh₃)₄ (1.6 g, 1.4 mmol, 0.1 eq.), Et₃N (17 g, 168 mmol, 16 eq.)in DMF (100 mL) was added formic acid (3.48 g, 75.6 mmol, 5.4 eq.)dropwise over 5 min. The mixture was warmed to 110° C. over 2 h. Then,the reaction mixture was diluted (water), extracted (ethyl acetate),washed (brine), dried (Na₂SO₄), filtered and evaporated to dryness togive the crude product, which was purified by silica gel columnchromatography (pet. ether:ethyl acetate, 1:1) to give7-hydroxyquinoline-3-carbaldehyde as yellow solid (1.45 g, yield 60%).ESI-MS: 216 (M+H)⁺.

To a solution of 7-hydroxyquinoline-3-carbaldehyde (1.3 g, 7.5 mmol) inDMF (30 mL) was added 1-bromo-heptane (5.37 g, 30 mmol, 4 eq.) and K₂CO₃(2.0 g, 15 mmol, 2 eq.). The mixture was warmed to 60° C. over 3 h.Then, the reaction mixture was diluted (water), extracted (ethylacetate), washed (brine), dried (Na₂SO₄), filtered and evaporated todryness to give the crude product, which was purified by silica gelcolumn chromatography (pet. ether:ethyl acetate, 10:1) to give7-(heptyloxy)quinoline-3-carbaldehyde as yellow solid (610 mg, yield30%). ESI-MS: 272 (M+H)⁺.

To a solution of 7-(heptyloxy)quinoline-3-carbaldehyde (1.6 g, 5.9 mmol)in THF (30 mL) at 0° C. was added CH₃MgI (2 g, 12 mmol, 2 eq.) dropwiseover 10 min. The mixture was warmed to it over 8 h. Then, the reactionmixture was quenched (water), extracted (ethyl acetate), washed (brine),dried (Na₂SO₄), filtered and evaporated to dryness to give the crudeproduct, which was purified by silica gel column chromatography (pet.ether:ethyl acetate, 1:5) to give 1-(7-(heptyloxy)quinolin-3-yl)ethanolas yellow oil (1.35 g, yield 80%). ESI-MS: 288 (M+H)⁺.

To a solution of oxalyl chloride (987 mg, 7.8 mmol, 1.5 eq.) in dryCH₂Cl₂ (40 mL) was added slowly DMSO (1.6 g, 20.8 mmol, 4 eq.) at −78°C. under N₂ After 30 min, 1-(7-(heptyloxy)quinolin-3-yl)ethanol (1.5 g,5.2 mmol) was added dropwise at −78° C. The mixture was stirred for 2 hat −78° C., and then Et₃N (3.2 g, 31 mmol, 6 eq.) was added at −78° C.After 20 min, the mixture was warmed to room temperature. Then thereaction mixture was added water (30 mL), extracted (DCM), washed(brine), dried (Na₂SO₄), filtered and evaporated to dryness to give thecrude product, which was purified by silica gel column chromatography(PE-EA, 10:1) to give 1-(7-(heptyloxy)quinolin-3-yl)ethanone as yellowsolid (1.08 g, yield 73%). ESI-MS: 286 (M+H)⁺.

A mixture of 1-(7-(heptyloxy)quinolin-3-yl)ethanone (730 mg, 2.56 mmol),EtOH (2 mL), H₂O (3 mL), (NH₄)₂ CO₃ (1.47 g, 15.36 mmol, 6 eq.), andNaCN (251 mg, 5.12 mmol, 2 eq.) was stirred for 16 h at 60° C. Then, thereaction mixture was added water (20 mL), extracted (EA), washed(brine), dried (Na₂SO₄), filtered and evaporated to dryness to give thecrude product, which was purified by silica gel column chromatography(PE-EA, 1:2) to give5-(7-(heptyloxy)quinolin-3-yl)-5-methylimidazolidine-2,4-dione as yellowsolid (480 mg, yield 53%). ESI-MS: 356 (M+H)⁺.

A mixture of5-(7-(heptyloxy)quinolin-3-yl)-5-methylimidazolidine-2,4-dione (1 g, 2.8mmol), EtOH (2 mL), H₂O (4 mL), and NaOH (2.24 g, 56mmol, 20 eq.) wasstirred for 4 day at is 110° C. Then, the reaction mixture was added HCl(40%) to adjust its pH=5 and then2-amino-2-(7-(heptyloxy)quinolin-3-yl)propanoic acid was generated asyellow solid (740 mg, yield 80%), without further purification for nextstep. ESI-MS: 331 (M+H)⁺.

To a solution of crude 2-amino-2-(7-(heptyloxy)quinolin-3-yl)propanoicacid (600 mg, 1.8 mmol) in dry THF (30 mL) was added LAH (138 mg, 3.6mmol, 2 eq.) at 0° C. under N₂. Then, the mixture was warmed to roomtemperature for 3 h. Then the reaction mixture was added water (1 mL),diluted (EA), filtered and dried (Na₂SO₄), evaporated to dryness to givethe crude product, which was purified by prep-HPLC to give2-amino-2-(7-(heptyloxy)quinolin-3-yl)propan-1-ol as white solid (200mg, yield 35%). ESI-MS: 317 (M+H)+.

Example 14 6-(trans-4-tert-Butyl-cyclohexyloxy)-quinoline

Quinolin-6-ol (7.52 g, 0.0518 mol, Aldrich), 4-tert-butyl-cyclohexanol(9.71 g, 0.0621 mol) and triphenylphosphine (16.32 g, 0.06222 mol,Aldrich) were placed in a flask, and dissolved in tetrahydrofuran (150mL, Acros). The reaction was cooled in a cold water bath. Diisopropylazodicarboxylate (13.0 mL, 0.0620 mol, Acros) in tetrahydrofuran (50 mL,Acros) was then added dropwise. The reaction mixture was then allowed tostir at room temperature. After 26 h,the solvent was removed, and theresidue was taken up in DCM. Silica gel was added and the solvent wasremoved. The residue was then purified via silica gel chromatographyusing 0-40% ethyl acetate in hexanes as eluent to give the product(Rf=0.22 in 3:1 hexanes/ethyl acetate), 6.11 g yield (42%).

Example 15 6-(trans-4-tert-butyl-cyclohexyloxy)-quinoline 1-oxide

6-(trans-4-tert-Butyl-cyclohexyloxy)-quinoline (6.11 g, 0.0216 mol) wasdissolved in acetone (100 mL, Acros), followed by m-chloroperbenzoicacid (11.7 g, 0.0542 mol, Aldrich) in small portions at RT. The reactionmixture was then stirred at 23° C. for 1.5 hours. Sodium thiosulfate wasadded, 97.6 mL of a saturated aqueous solution. The reaction mixture wasthen allowed to stir for 45 minutes. Excess of solvents were removedunder vacuum, and the residue was diluted with water and extracted withmethylene chloride. Combined organic layers were treated with magnesiumsulfate, filtered and evaporated. The residue was taken up in methylenechloride and silica gel was added. The solvents were removed. Theresidue was then purified via chromatography (SiO₂, 0-10% MeOH/DCM) togive the product (Rf=0.19 in 5% methanol/methylene chloride). Theappropriate fractions were combined to give the product in 4.99 g yield(77%).

Example 162-[6-(trans-4-tert-butyl-cyclohexyloxy)-quinolin-2-yl]-2-nitro-propionicacid ethyl ester

A solution of ethyl 2-nitropropionate (5.94 g, 0.0404 mol, Aldrich) inN,N-dimethylformamide (40 mL, Acros) was added to6-(trans-4-tert-butyl-cyclohexyloxy)-quinoline 1-oxide (10.08 g, 0.03367mol) and acetic anhydride (4.765 mL, 0.05050 mol, Aldrich) inN,N-dimethylformamide (80 mL, Aldrich) dropwise. The reaction mixturewas then stirred at RT for 3 d. The reaction was partitioned betweenNa₂CO₃ (sat), and DCM. Organic layer was collected, and dried overMgSO₄. The drying agent was filtered and the solvent was removed byevaporation. The mixture was then purified via chromatography (SiO₂,0-5% methanol in methylene chloride as eluent) to give the product(solvent front in 5% methanol/methylene chloride, Rf=0.72 in 3:1hexanes/ethyl acetate). Appropriate fractions were combined andevaporated to give 7.37 g of the product as a white solid (51%).

Example 172-Amino-2-[6-(trans-4-tert-butyl-cyclohexyloxy)-quinolin-2-yl]-propionicacid ethyl ester

2-[6-(trans-4-tert-Butyl-cyclohexyloxy)-quinolin-2-yl]-2-nitro-propionicacid ethyl ester (76 mg, 0.00018 mol) was dissolved in acetic acid (1.0mL, Fisher), and cooled in a 20° C. water bath, followed by addition ofzinc (115 mg, 0.00176 mol, Aldrich) in small portions. After 2 h, thereaction was diluted with acetic acid and acetonitrile and filteredthrough a PTFE filter. The excess of solvents were removed under vacuum,and the residue was purified via preparative HPLC (10-90% acetonitrilewith 0.1% TFA) to give a pure product (18 mg, 20%) as TFA salt.

Example 182-Amino-2-[6-(trans-4-tert-butyl-cyclohexyloxy)-quinolin-2-yl]-propan-1-ol

2-Amino-2-[6-(trans-4-tert-butyl-cyclohexyloxy)-quinolin-2-yl]-propionicacid ethyl ester (18 mg as TFA salt, 0.000035 mol) was dissolved in amixture of methanol (2.0 mL, Acros) and tetrahydrofuran (2.0 mL, Acros),then sodium tetrahydroborate (3.8 mg, 0.00010 mol, Aldrich) was added.After 2 h, additional sodium tetrahydroborate (8.1 mg, 0.00021 mol,Aldrich) was added. After an additional 2 h, the reaction mixture wasquenched with 250 uL of to saturated NH₄Cl, and solvents were removedunder vacuum. DMSO (½ mL) was added to the residue but it did notcompletely solubilize. The material was partitioned between methylenechloride and water. The emulsion was broken up by adding saturatedsodium chloride and solid sodium chloride. The organics were dried withmagnesium sulfate, filtered and evaporated. The residue was purified viapreparative HPLC to give pure product (6.3 mg, 38%) as TFA salt. MS:m/z=357.20 [M+H]+. ¹H NMR (400 MHz, CHLOROFORM-d) δ ppm: 8.19 (d, J=8.5Hz, 1H), 7.96 (d, J=9.3 Hz, 1H), 7.39-7.46 (m, 2H), 7.13 (d, J=2.8 Hz,1H), 4.24-4.36 (m, 1H), 4.12-4.21 (m, 2H), 2.23-2.32 (m, 2H), 1.88-1.96(m, 2H), 1.78 (s, 3H), 1.41-1.54 (m, 2H), 1.10-1.27 (m, 3H), 0.91 (s,9H).

Example 192-[6-(trans-4-tert-Butyl-cyclohexyloxy)-5-iodo-quinolin-2-yl]-2-nitro-propionicacid ethyl ester

N-Iodosuccinimide (3446 mg, 0.01532 mol, Aldrich) and zirconiumtetrachloride (481 mg, 0.00206 mol, Aldrich) were added to a solution of2-[6-(trans-4-tert-Butyl-cyclohexyloxy)-quinolin-2-yl]-2-nitro-propionicacid ethyl ester (4.37 g, 0.0102 mol) in methylene chloride (150 mL,Acros) and was stirred at room temperature for 1 h. The reaction wasfiltered through Celite, washed, then filtered through a paper filter.The solution was held at room temperature for 2 h. The solvent was thenevaporated to approximately 100 mL and then silica gel was added. Thesolvent was removed under reduced pressure, and the residue was purifiedby silica gel chromatography using 0-25% ethyl acetate in hexanes aseluent to give the product (Rf=0.33 in 7:1 hexanes/ethyl acetate) in5.28 g yield (93%). Note that the product contains 6% of an unidentifiedmaterial (possibly the analogous chloride).

Example 19A2-[6-(trans-4-tert-butyl-cyclohexyloxy)-5-trifluoromethyl-quinolin-2-yl]-2-nitro-propionicacid ethyl ester

A solution of2-[6-(trans-4-tert-Butyl-cyclohexyloxy)-5-iodo-quinolin-2-yl]-2-nitro-propionicacid ethyl ester (5.28 g, 9.52 mmol), hexamethylphosphoramide (8.37 mL,47.6 mmol, Fluka) in N,N-dimethylformamide (55 mL, Aldrich) was degassedby stirring vigorously while forcing argon through the apparatus (5min). To this was added copper(I)iodide (3.26 g, 17.1 mmol, Aldrich) andmethyl fluorosulphonyldifluoroacetate (6.24 mL, 47.6 mmol, Aldrich) andthe reaction was stirred in a sealed vessel at 80° C. under anatmosphere of argon. After stirring for 17 hours, the reaction wasevaporated, then diluted with methylene chloride. Silica to gel wasadded and the solvent removed under reduced pressure. The material waspurified by silica gel chromatography using 0-35% ethyl acetate inhexanes as eluent (Rf=0.25 in 7:1 ethyl acetate/hexanes) to give theproduct in 4.71 g (100%) yield. The impurity seen in2-[6-(trans-4-tert-Butyl-cyclohexyloxy)-5-iodo-quinolin-2-yl]-2-nitro-propionicacid ethyl ester is still present, in less than 5%.

Example 202-Amino-2-[6-(trans-4-tert-butyl-cyclohexyloxy)-5-trifluoromethyl-quinolin-2-yl]-propionicacid ethyl ester

2-Amino-2-[6-(trans-4-tert-butyl-cyclohexyloxy)-5-trifluoromethyl-quinolin-2-yl]-propionicacid ethyl ester was synthesized as per2-amino-2-[6-(trans-4-tert-butyl-cyclohexyloxy)-quinolin-2-yl]-propionicacid ethyl ester (Example 17) in 20% yield (TFA salt) using2-[6-(trans-4-tert-Butyl-cyclohexyloxy)-5-trifluoromethyl-quinolin-2-yl]-2-nitro-propionicacid ethyl ester as starting material.

Example 212-Amino-2-[6-(trans-4-tert-butyl-cyclohexyloxy)-5-trifluoromethyl-quinolin-2-yl]-propan-1-ol

2-Amino-2-[6-(trans-4-tert-butyl-cyclohexyloxy)-5-trifluoromethyl-quinolin-2-yl]-propan-1-ol was synthesized as per2-amino-2-[6-(trans-4-tert-butyl-cyclohexyloxy)-quinolin-2-yl]-propan-1-ol(Example 18) in 49% yield (TFA salt) using2-amino-2-[6-(trans-4-tert-butyl-cyclohexyloxy)-5-trifluoromethyl-quinolin-2-yl]-propionicacid ethyl ester as starting material (TFA salt). MS: m/z=425.20 [M+H]+.¹H NMR (400 MHz, CHLOROFORM-d) δ ppm: 8.70 (d, J=8.8 Hz, 1H), 8.15 (d,J=9.5 Hz, 1H), 7.62 (d, J=9.5 Hz, 1H), 7.50 (d, J=9.3 Hz, 1H), 4.33-4.45(m, 1H), 4.08-4.25 (m, 2H), 2.15-2.27 (m, 2H), 1.85-1.98 (m, 2H), 1.78(s, 3H), 1.48-1.64 (m, 2H), 1.04-1.24 (m, 3H), 0.90 (s, 9H).

Example 222-Amino-2-[6-(trans-4-tert-butyl-cyclohexyloxy)-quinolin-2-yl]-propionicacid ethyl ester Enantiomer 1 and2-Amino-2-[6-(trans-4-tert-butyl-cyclohexyloxy)-quinolin-2-yl]-propionicacid ethyl ester Enantiomer 2

2-Amino-2-[6-(trans-4-tert-butyl-cyclohexyloxy)-quinolin-2-yl]-propionicacid ethyl ester (racemic mixture, 522 mg) was separated by chiralchromatography (ChiralPak IC (3×15 cm) 35% EtOH (0.1% DEA)/CO₂ 100 bar75 mL/min 220 nm inj vol 0.5 mL 26 mg/mL methanol). Isolated were 220 mg(42%) of enantiomer 1 (RT=1.65 min on Chiralpak IC 15×0.46 cm, 40%ethanol(DEA)/CO2, 100 bar, 3 ml/min, 220 mu) and 226 mg (43%) ofenantiomer 2 (RT=2.25 min, same conditions).

Example 232-Amino-2-[6-(trans-4-tert-butyl-cyclohexyloxy)-quinolin-2-yl]-propan-1-olEnantiomer 1

2-Amino-2-[6-(trans-4-tert-butyl-cyclohexyloxy)-quinolin-2-yl]-propan-1-olEnantiomer 1 was synthesized as per2-Amino-2-[6-(trans-4-tert-butyl-cyclohexyloxy)-quinolin-2-yl]-propan-1-ol(Example 18) in 31% yield as bis-TFA salt, using2-amino-2-[6-(trans-4-tert-butyl-cyclohexyloxy)-quinolin-2-yl]-propionicacid ethyl ester Enantiomer 1 as starting material. MS: m/z=357.30[M+H]+. ¹H NMR (400 MHz, CHLOROFORM-d) δ ppm: 8.18 (d, J=8.5 Hz, 1H),7.96 (d, J=9.0 Hz, 1H), 7.38-7.47 (m, 2H), 7.13 (d, J=2.8 Hz, 1H),4.25-4.35 (m, 1H), 4.11-4.23 (m, 2H), 2.24-2.32 (m, 2H), 1.88-1.97 (m,2H), 1.83 (s, 3H), 1.41-1.54 (m, 2H), 1.10-1.31 (m, 3H), 0.91 (s, 9H).

Example 242-Amino-2-[6-(trans-4-tert-butyl-cyclohexyloxy)-quinolin-2-yl]-propan-1-olEnantiomer 2

2-Amino-2-[6-(trans-4-tert-butyl-cyclohexyloxy)-quinolin-2-yl]-propan-1-olEnantiomer 2 was synthesized as per2-amino-2-[6-(trans-4-tert-butyl-cyclohexyloxy)-quinolin-2-yl]-propan-1-ol(Example 18) in 19% yield as bis-TFA salt, using2-amino-2-[6-(trans-4-tert-butyl-cyclohexyloxy)-quinolin-2-yl]-propionicacid ethyl ester Enantiomer 2 as starting material. MS: m/z=357.20[M+H]+. ¹HNMR (400 MHz, CHLOROFORM-d) δ ppm: 8.19 (d, J=8.5 Hz, 1H),7.96 (d, J=9.3 Hz, 1H), 7.38-7.48 (m, 2H), 7.13 (d, J=2.8 Hz, 1H),4.25-4.35 (m, 1H), 4.10-4.23 (m, 2H), 2.23-2.32 (m, 2H), 1.88-1.98 (m,2H), 1.82 (s, 3H), 1.41-1.54 (m, 2H), 1.10-1.28 (m, 3H), 0.91 (s, 9H).

Example 252-Amino-2-[6-(trans-4-tert-butyl-cyclohexyloxy)-5-trifluoromethyl-quinolin-2-yl]-propionicacid ethyl ester Enantiomer 1 and2-Amino-2-[6-(trans-4-tert-butyl-cyclohexyloxy)-5-trifluoromethyl-quinolin-2-yl]-propionicacid ethyl ester Enantiomer 2

Racemic2-Amino-2-[6-(trans-4-tert-butyl-cyclohexyloxy)-5-trifluoromethyl-quinolin-2-yl]-propionicacid ethyl ester (755 mg) was separated into two enantiomers byChiralpak IC (3×15cm); 20% methanol (0.1% DEA)/CO₂, 100 bar; 75 mL/min,220 nm; inj vol. 0.3 mL, 38 mg/mL methanol. Isolated were: Enantiomer 1:323 mg (43%), >99%, >99% ee RT=1.22 min on Chiralpak IC (15×0.46 cm);40% methanol (DEA)/CO₂, 100 bar; 3 mL/min, 220 nm Enantiomer 2: 322 mg(43%), >99%, >98% ee RT=1.34 min on Chiralpak IC (15×0.46 cm); 40%methanol (DEA)/CO₂, 100 bar; 3 mL/min, 220 nm

Example 262-Amino-2-[6-(trans-4-tert-butyl-cyclohexyloxy)-5-trifluoromethyl-quinolin-2-yl]-propan-1-olEnantiomer 1

2-Amino-2-[6-(trans-4-tert-butyl-cyclohexyloxy)-5-trifluoromethyl-quinolin-2-yl]-propan-1-olEnantiomer 1 was synthesized as per2-amino-2-[6-(trans-4-tert-butyl-cyclohexyloxy)-quinolin-2-yl]-propan-1-ol(Example 18) in 52% yield as bis-TFA salt using2-amino-2-[6-(trans-4-tert-butyl-cyclohexyloxy)-5-trifluoromethyl-quinolin-2-yl]-propionicacid ethyl ester Enantiomer 1 as starting material. MS: m/z=425.30[M+H]+. ¹NMR (400 MHz, CHLOROFORM-d) δ ppm: 8.68 (d, J=8.8 Hz, 1H), 8.12(d, J=9.0 Hz, 1H), 7.47-7.60 (m, 2H), 4.31-4.44 (m, 1H), 4.09-4.26 (m,2H), 2.13-2.25 (m, 2H), 1.85-1.95 (m, 2H), 1.81 (s, 3H), 1.48-1.62 (m,2H), 1.03-1.22 (m, 3H), 0.90 (s, 9H).

Example 272-Amino-2-[6-(trans-4-tert-butyl-cyclohexyloxy)-5-trifluoromethyl-quinolin-2-yl]-propan-1-olEnantiomer 2

2-Amino-2-[6-(trans-4-tert-butyl-cyclohexyloxy)-5-trifluoromethyl-quinolin-2-yl]-propan-1-olEnantiomer 2 was synthesized as per2-amino-2-[6-(trans-4-tert-butyl-cyclohexyloxy)-quinolin-2-yl]-propan-1-ol(Example 17) in 55% yield as bis-TFA salt using2-amino-2-[6-(trans-4-tert-butyl-cyclohexyloxy)-5-trifluoromethyl-quinolin-2-yl]-propionicacid ethyl ester Enantiomer 2 as starting material. MS: m/z=425.30[M+H]+. ¹H NMR (400 MHz, CHLOROFORM-d) δ ppm: 8.68 (d, J=8.5 Hz, 1H),8.13 (d, J=9.5 Hz, 1H), 7.56 (d, J=9.8 Hz, 1H), 7.51 (d, J=9.0 Hz, 1H),4.30-4.44 (m, 1H), 4.06-4.24 (m, 2H), 2.12-2.26 (m, 2H), 1.85-1.95 (m,2H), 1.80 (s, 3H), 1.48-1.62 (m, 2H), 1.03-1.22 (m, 3H), 0.90 (s, 9H).

Example 28{1-[6-(trans-4-tert-butyl-cyclohexyloxy)-quinolin-2-yl]-2-hydroxy-1-methyl-ethyl}-carbamicacid tert-butyl ester Enantiomer 1

2-Amino-2-[6-(trans-4-tert-butyl-cyclohexyloxy)-quinolin-2-yl]-propan-1-olEnantiomer 1 (0.1055 g, 0.0001805 mol as bis-TFA salt) was dissolved inchloroform (2.0 mL, Aldrich) along with di-tert-Butyldicarbonate (89 mg,0.00041 mol, Aldrich). Saturated aqueous sodium bicarbonate solution(1.0 mL, 0.010 mol) was added and the reaction was stirred vigorouslyfor 16 h. The reaction was diluted with methylene chloride and theaqueous layer removed. Drying with MgSO₄, filtering, evaporation ofsolvent and purification by silica gel chromatography using 0-50% ethylacetate in hexanes as eluent to give the product (Rf=0.20 in 3:1hexanes/ethyl acetate) in 81 mg yield (98%).

Example 29[1-[6-(4-trans-tert-butyl-cyclohexyloxy)-quinolin-2-yl]-2-(di-tert-butoxy-phosphoryloxy)-1-methyl-ethyl]-carbamicacid tert-butyl ester Enantiomer 1

To a solution of{1-[6-(trans-4-tert-Butyl-cyclohexyloxy)-quinolin-2-yl]-2-hydroxy-1-methyl-ethyl}-carbamicacid tert-butyl ester Enantiomer 1 (81.0 mg, 0.000177 mol) and1H-tetrazole (0.124 g, 0.00177 mol, Waterstone) in tetrahydrofuran (1.2mL, Acros) was added di-tert-butyl N,N-diethylphosphoramidite (247 uL,0.000887 mol, Aldrich) at rt. After 2 h, the phosphite was oxidized withhydrogen peroxide (118 uL, 0.00115 mol, Aldrich) and stirred for 1 hour.The reaction was then quenched with 10% NaS₂O₃ in saturated sodiumbicarbonate, extracted with EtOAc, washed with saturated sodium chlorideand then dried with Na₂SO₄. The drying agent was filtered and theorganic layer was concentrated in vacuo, yielding the crude product. Thecrude was taken up in DCM and purified using silica gel chromatographyusing 0-100% ethyl acetate in hexanes as eluent. Isolated was theproduct (Rf=0.47 in 1:1 hexanes/ethyl acetate) in 86 mg yield (75%).

Example 30 Phosphoric acidmono-{2-amino-2-[6-(4-tert-butyl-cyclohexyloxy)-quinolin-2-yl]-propyl}ester Enantiomer 1

To[1-[6-(trans-4-tert-Butyl-cyclohexyloxy)-quinolin-2-yl]-2-(di-tert-butoxy-phosphoryloxy)-1-methyl-ethyl]carbamicacid tert-butyl ester Enantiomer 1 (86 mg, 0.00013 mol) was added 12 Mhydrogen chloride in water (1.0 mL, Fisher) and acetic acid (5.0 mL,Fisher) and the solution was stirred for 1.5 h at RT. Removal of solventgave an oil, which was purified by preparative HPLC to give the productin 14 mg yield as bis-TFA salt (16%). MS: m/z=437.62 [M+H]+. ¹H NMR (400MHz, METHANOL-d₄) δ ppm: 8.30 (d, J=8.5 Hz, 1H), 8.00 (d, J=9.3 Hz, 1H),7.63 (d, J=8.3 Hz, 1H), 7.40 (dd, J=9.2, 2.6 Hz, 1H), 7.33 (d, J=2.5 Hz,1H), 4.35-4.45 (m, 1H), 4.27-4.35 (m, 1H), 3.95-4.01 (m, 1H), 2.24-2.34(m, 2H), 1.89-1.98 (m, 2H), 1.79 (s, 3H), 1.38-1.53 (m, 2H), 1.22-1.37(m, 2H), 1.08-1.20 (m, J=13.6 Hz, 1H), 0.94 (s, 9H).

Example 31{1-[6-(trans-4-tert-Butyl-cyclohexyloxy)-5-trifluoromethyl-quinolin-2-yl]-2-hydroxy-1-methyl-ethyl}-carbamicacid tert-butyl ester Enantiomer 1

{1-[6-(trans-4-tert-Butyl-cyclohexyloxy)-5-trifluoromethyl-quinolin-2-yl]-2-hydroxy-1-methyl-ethyl}-carbamicacid tert-butyl ester Enantiomer 1 was synthesized as per{1-[6-(trans-4-tert-Butyl-cyclohexyloxy)-quinolin-2-yl]-2-hydroxy-1-methyl-ethyl}-carbamicacid tert-butyl ester Enantiomer 1 (Example 28) in 95% yield using2-amino-2-[6-(trans-4-tert-butyl-cyclohexyloxy)-5-trifluoromethyl-quinolin-2-yl]-propan-1-olEnantiomer 1 as starting material (bis-TFA salt).

Example 32[1-[6-(trans-4-tert-Butyl-cyclohexyloxy)-5-trifluoromethyl-quinolin-2-yl]-2-(di-tert-butoxy-phosphoryloxy)-1-methyl-ethyl]-carbamicacid tert-butyl ester Enantiomer 1

[1-[6-(trans-4-tert-Butyl-cyclohexyloxy)-5-trifluoromethyl-quinolin-2-yl]-2-(di-tert-butoxy-phosphoryloxy)-1-methyl-ethyl]-carbamicacid tert-butyl ester Enantiomer 1 was synthesized as per[1-[6-(trans-4-tert-Butyl-cyclohexyloxy)-quinolin-2-yl]-2-(di-tert-butoxy-phosphoryloxy)-1-methyl-ethyl]-carbamicacid tert-butyl ester Enantiomer 1 (Example 29) in 68% yield using{1-[6-(trans-4-tert-Butyl-cyclohexyloxy)-5-trifluoromethyl-quinolin-2-yl]-2-hydroxy-1-methyl-ethyl}-carbamicacid tert-butyl ester Enantiomer 1 as starting material.

Example 33 Phosphoric acidmono-{2-amino-2-[6-(trans-4-tert-butyl-cyclohexyloxy)-5-trifluoromethyl-quinolin-2-yl]-propyl}esterEnantiomer 1

Phosphoric acidmono-{2-amino-2-[6-(trans-4-tert-butyl-cyclohexyloxy)-5-trifluoromethyl-quinolin-2-yl]-propyl}ester Enantiomer 1 was synthesized as per Phosphoric acidmono-{2-amino-2-[6-(trans-4-tert-butyl-cyclohexyloxy)-quinolin-2-yl]-propyl}esterEnantiomer 1 (Example 30) in 88% yield as bis-HCl salt using[1-[6-(trans-4-tert-Butyl-cyclohexyloxy)-5-trifluoromethyl-quinolin-2-yl]-2-(di-tert-butoxy-phosphoryloxy)-1-methyl-ethyl]-carbamicacid tert-butyl ester Enantiomer 1 as starting material. (Note that HPLCpurification was not necessary and compound was isolated as bis-HClsalt). MS: m/z=505.20 [M+H]+. ¹H NMR (400 MHz, METHANOL-d₄) δ ppm: 8.67(d, J=8.5 Hz, 1H), 8.31 (d, J=9.5 Hz, 1H), 7.85 (d, J=9.3 Hz, 1H), 7.79(d, J=9.3 Hz, 1H), 4.50-4.62 (m, 1H), 4.34-4.42 (m, 1H), 4.26-4.34 (m,1H), 2.17-2.29 (m, 2H), 1.88-1.98 (m, 2H), 1.80 (s, 3H), 1.45-1.59 (m,2H), 1.20-1.34 (m, 2H), 1.07-1.19 (m, 1H), 0.93 (s, 9H).

Example 34 6-Heptyloxyquinoline

Quinolin-6-ol (1, 2.00 g, 0.0138 mol), 1-heptanol (1.60 g, 0.0138 mol)and triphenylphosphine (4.34 g, 0.0165 mol) were placed in a flask, anddissolved in tetrahydrofuran (200 mL, 2 mol) at 23° C. A solution ofdiisopropyl azodicarboxylate (3.46 mL, 0.0165 mol) in THF (5 mL) wasthen added dropwise. The reaction mixture was then allowed to stir at23° C. for 15 hours. Solvent was removed, and the residue was thenpurified via chromatography (SiO₂, 120 g, 5% MeOH in DCM) to give 2.95 gproduct (88%). ¹H NMR (CHLOROFORM-d) δ: 8.76 (dd, J=4.0, 1.5 Hz, 1H),8.02 (dd, J=15.9, 8.7 Hz, 2H), 7.31-7.44 (m, 2H), 7.07 (d, J=2.8 Hz,1H), 4.09 (t, J=6.5 Hz, 2H), 1.79-1.96 (m, 2H), 1.45-1.60 (m, 4H),1.14-1.45 (m, 4H), 0.82-0.99 (m, 3H). MS (M+1): 244.20.

Example 35 6-Heptyloxyquinoline 1-oxide

6-Heptyloxyquinoline (1000.0 mg, 0.0041094 mol) was dissolved in acetone(20 mL, 0.3 mol), followed by m-chloroperbenzoic acid (1060 mg, 0.00493mol) in small portions at 23° C. The reaction mixture was then stirredat 23° C. for 3 hours, and LC/MS indicated all the SM was converted tothe desired product. A solution of sodium thiosulfate (1950 mg, 0.0123mol) was added. The reaction mixture was then allowed to stir for 1hour. Excess of solvents were removed under vacuum, and the residue wastreated with DCM (100×3 mL). Combined organic layers were treated withsodium sulfate and filtrated. The crude mixture was then purified viachromatography (SiO₂, 120g, 0-8% MeOH/DCM) to give 653 mg (53%) pureproduct. ¹H NMR (CHLOROFORM-d) δ: 8.65 (d, J=9.5 Hz, 1H), 8.38 (d, J=5.8Hz, 1H), 7.60 (d, J=8.5 Hz, 1H), 7.38 (dd, J=9.5, 2.5 Hz, 1H), 7.23 (dd,J=8.3, 6.0 Hz, 1H), 7.09 (d, J=2.5 Hz, 1H), 4.08 (t, J=6.5 Hz, 2H),1.80-1.92 (m, 2H), 1.44-1.57 (m, 2H), 1.28-1.44 (m, 6H), 0.90 (t, J=6.8Hz, 3H). MS (M+1): 260.20.

Example 36 Ethyl 2-(6-heptyloxyquinolin-2-yl)-2-nitropropanoate

A solution of 6-Heptyloxy-quinoline 1-oxide (500 mg, 0.002 mol) andacetic anhydride (0.255 mL, 0.00270 mol) in N,N-dimethylformamide (5 mL,0.06 mol) was added to a solution of ethyl 2-nitropropionate (284 mg,0.00193 mol) in DMF (2 mL) at at 23° C. dropwise. The reaction mixturewas then stirred at 23° C. for 12 hours. The reaction mixture was thenheated at 50° C. for 6 hours. Solvent was removed under vacuum, and theresidue was treated with Na₂CO₃ (sat), and DCM. Organic layer wascollected, and dried over Na₂SO₄. The mixture was then purified viachromatography (SiO₂, 24 g, ethyl acetate/hexanes 0-35%) to give a pureproduct 523 mg (70%). ¹H NMR (CHLOROFORM-d) δ: 8.09 (d, J=8.5 Hz, 1H),7.95 (d, J=9.0 Hz, 1H), 7.52 (d, J=8.8 Hz, 1H), 7.39 (dd, J=9.3, 2.5 Hz,1H), 7.08 (d, J=2.5 Hz, 1H), 4.40 (q, J=7.0 Hz, 2H), 4.09 (t, J=6.5 Hz,2H), 2.40 (s, 3H), 1.82-1.93 (m, 2H), 1.45-1.56 (m, 2H), 1.26-1.44 (m,9H), 0.92 (t, J=6.7 Hz, 3H). MS (M+1): 389.30.

Example 37 Ethyl 2-amino-2-(6-heptyloxyquinolin-2-yl)propanoate

Ethyl 2-(6-heptyloxyquinolin-2-yl)-2-nitropropanoate (325.0 mg,0.0008366 mol) was dissolved in acetic acid (5 mL, 0.09 mol), and cooledto at 10° C., followed by zinc (547.1 mg, 0.008366 mol;) in very smallportions to keep the temperature constant. The reaction mixture was thenallowed to stir for additional for 5 hours, and filtered through aCelite pad. The excess is of solvents were removed under vacuum, and theresidue was purified via HPLC (10-100% acetonitrile with 0.1% formicacid) to give a pure product (57.0 mg, 14%). ¹H NMR (CHLOROFORM-d) δ:8.05 (d, J=8.5 Hz, 1H), 7.94 (d, J=9.3 Hz, 1H), 7.57 (d, J=8.5 Hz, 1H),7.36 (dd, J=9.3, 2.5 Hz, 1H), 7.06 (d, J=2.5 Hz, 1H), 4.20 (q, J=7.0 Hz,2H), 4.07 (t, J=6.5 Hz, 2H), 1.77-1.93 (m, 5H), 1.44-1.57 (m, 2H),1.28-1.44 (m, 6H), 1.22 (t, J=7.2 Hz, 3H), 0.90 (t, J=6.8 Hz, 3H). MS(M+1): 359.30.

Example 38 2-amino-2-(6-(heptyloxy)quinolin-2-yl)propan-1-ol

Ethyl 2-amino-2-(6-heptyloxyquinolin-2-yl)propanoate (7, 5.0 mg,0.000014 mol) was dissolved in a mixture of methanol (1.0 mL, 0.025 mol)and tetrahydrofuran (1.0 mL, 0.012 mol), followed by sodiumtetrahydroborate (1.06 mg, 0.0000279 mol) in small portions at 23° C.The reaction mixture was then allowed to stir for 2 hours. The reactionmixture was quenched with NH₄Cl (sat), and solvents were removed undervacuum. The solid residue was then treated with MeOH/DCM, and the crudeproduct was purified via HPLC to give pure product (4 mg, 70%). ¹H NMR(CHLOROFORM-d) δ: 8.22 (d, J=8.5 Hz, 1H), 7.97 (d, J=9.3 Hz, 1H),7.39-7.51 (m, 2H), 7.10 (d, J=2.5 Hz, 1H), 4.14 (br. s., 2H), 4.08 (t,J=6.5 Hz, 2H), 1.81-1.93 (m, 2H), 1.77 (s, 3H), 1.50 (d, J=8.0 Hz, 2H),1.21-1.44 (m, 6H), 0.81-0.99 (m, 3H). MS (ES, M+1): 317.30.

Example 39 5-(Heptyloxy)-2-nitrobenzaldehyde

5-Hydroxy-2-nitrobenzaldehyde (3.60 g, 0.0215 mol), 1-heptanol (2.50 g,0.0215 mol) and triphenylphosphine polymer bound (3 mmol/g loading; 9 g,0.028 mol) were placed in a flask followed by tetrahydrofuran (300 mL, 4mol). A solution of azodicarboxylic acid di-tert-butyl ester (5.94 g,0.0258 mol) in THF (15 mL) was then added dropwise at 23° C. Thereaction mixture was then allowed to stir for 1 day. Filtered through acelite pad, and the crude mixture was then purified via chromatography(SiO₂, 120 g, 0-100% ethyl acetate/hexanes) to give a pure product (3.25g, 57%). ¹H NMR (CHLOROFORM-d) 8: 10.50 (s, 1H), 8.16 (d, J=9.0 Hz, 1H),7.32 (d, J=3.0 Hz, 1H), 7.14 (dd, J=9.0, 2.8 Hz, 1H), 4.11 (t, J=6.5 Hz,2H), 1.76-1.92 (m, 2H), 1.42-1.53 (m, 2H), 1.21-1.42 (m, 6H), 0.82-0.99(m, 3H). MS (M+1): 266.20.

Example 40 2-(5-Heptyloxy)-2-nitrophenyl-1,3-dioxolane

5-(heptyloxyl)-2-nitrobenzaldehyde (3, 1.00 g, 0.00377 mol) and1,2-Bis-trimethylsilanyloxy-ethane (0.924 mL, 0.00377 mol) were placedin a 40 mL vial, followed by methylene chloride (50 mL, 0.8 mol). Thereaction mixture was then cooled to −78° C., and trimethylsilyltrifluoromethanesulfonate (0.03 mL, 0.0002 mol) was added dropwise underN₂. The reaction mixture was allowed to stir for about for 3 hours at−78° C. The reaction mixture was then warmed up to 23° C., and thencooled down to −78° C. again. A solution of NaOH (4N) was added toquench the reaction, and organic layer was separated. The crude mixturewas then purified via chromatography (SiO₂, 24 g, 0-35%ethylacetate/hexanes) to give the desired product (1030mg, 88%). ¹H NMR(CHLOROFORM-d) δ: 8.04 (d, J=9.0 Hz, 1H), 7.29 (d, J=2.8 Hz, 1H), 6.91(dd, J=9.0, 2.8 Hz, 1H), 6.58 (s, 1H), 3.98-4.12 (m, 6H), 1.74-1.90 (m,2H), 1.42-1.53 (m, 2H), 1.24-1.41 (m, 6H), 0.83-0.98 (m, 3H). MS (M+1):310.20.

Example 41 2-(1,3-Dioxolan-2-yl)-4-heptyloxyaniline

2-(5-Heptyloxy)-2-nitrophenyl-1,3-dioxolane (4, 950.00 mg, 0.0030709mol), platinum dioxide (40 mg, 0.0002 mol) and sodium acetate trihydrate(30 mg, 0.0002 mol) were placed in a pressure flask, followed by ethylacetate (50 mL, 0.5 mol). The reaction mixture was then purged under N₂for at least 3 times, and H₂ was introduced (purged 3 times) andmaintained at 52 psi for 3 hours. The reaction mixture was thenfiltered, and solvent was removed to give a pure product (855 mg, 100%).¹H NMR (CHLOROFORM-d) δ: 6.95 (d, J=2.8 Hz, 1H), 6.76 (dd, J=8.5, 2.8Hz, 1H), 6.63 (d, J=8.5 Hz, 1H), 5.84 (s, 1H), 4.01-4.18 (m, 4H),3.81-3.95 (m, 2H), 1.67-1.81 (m, 2H), 1.56 (br. s., 2H), 1.38-1.50 (m,2H), 1.20-1.38 (m, 4H), 0.84-0.95 (m, 3H). MS (M+1): 280.20.

Example 42 (R)-Methyl 3-(methoxymethoxy)-2-methylpropanoate

(R)-3-Hydroxy-2-methyl-propionic acid methyl ester (8.60 g, 0.0728 mol)was dissolved in methylene chloride (200 mL, 3 mol), followed byN,N-diisopropylethylamine (25.4 mL, 0.146 mol). The reaction mixture wasthen cooled to −78° C., and chloromethyl methyl ether (9.48 mL, 0.0874mol) was added dropwise. The reaction mixture was then allowed to warmup gradually over for 5 hours to 23° C., and quenched with Na₂CO₃ (sat).Organic layer was separated, and washed with water, brine, and thendried over Na₂SO₄. The to crude mixture was purified via chromatography(SiO₂, 120g, 0-30% ethyl acetate/hexanes) to give 10.3 g clear liquidproduct (87%). ¹H NMR (CHLOROFORM-d) δ: 4.61 (s, 2H), 3.71 (m, 4H), 3.58(dd, J=9.5, 5.5 Hz, 1H), 3.35 (s, 3H), 2.77 (m, 1H), 1.19 (d, J=7.0 Hz,3H).

Example 43 (R)-3-(methoxymethoxy)-2-methylpropanoic acid

(R)-Methyl 3-(methoxymethoxy)-2-methylpropanoate (7, 5.00 g, 0.0308 mol)was is dissolved in methanol (50 mL, 1 mol). The solution was thencooled to −78° C. Sodium methoxide (5.26 g, 0.0925 mol), was addedgradually (in very small portions) followed by water (1.11 mL, 0.0616mol) at −78° C. The reaction mixture was then stirred at 23° C. for 12hours. An additional portion of NaOMe (95%, 1.0 g) was added to thereaction mixture followed by H₂O (0.8 mL). The reaction mixture wasallowed to stir at 23° C. for 4 hours. All the solvents were removedunder vacuum, and before quenched with concentrated HCl to pH ˜5. Excessof solvents were removed under vacuum. The residue was extracted withDCM (50×3 mL). The combined organic layers were treated with water,brine and dried over Na₂SO₄. Removal of solvent gave a liquid product(4.37 g, 96%). ¹H NMR (CHLOROFORM-d) δ: 4.64 (s, 2H), 3.69-3.82 (m, 1H),3.56-3.68 (m, 1H), 3.37 (s, 3H), 2.80 (m, 1H), 1.23 (d, J=7.3 Hz, 3H).

Example 44(R)—N-2-(1,3-Dioxolan-2-yl)-4-heptyloxyphenyl-3-methoxymethoxy-2-methylpropanamide

(R)-3-(methoxymethoxy)-2-methylpropanoic acid (1.27 g, 0.00859 mol) wasdissolved in methylene chloride (20 mL, 0.4 mol), and cooled to at −78°C., followed by oxalyl chloride (0.909 mL, 0.0107 mol) and catalyticamounts of N,N-dimethylformamide (0.006 mL, 0.00007 mol). The reactionmixture was allowed to warm up gradually until bubbles appeared. Thereaction mixture was then maintained at that temperature until bubblingceased. The reaction mixture was then cooled down again to at −78° C.and excess reagent and solvent were removed under vacuum. The cruderesidue was then re-dissolved in THF (5 mL), and added drop wise to asolution of 2-(1,3-dioxolan-2-yl)-4-heptyloxyaniline (2.00 g, 0.00716mol) and triethylamine (2.00 mL, 0.0143 mol) in tetrahydrofuran (60 mL,0.8 mol) at −78° C. The reaction mixture was then gradually warmed up toat 23° C. for additional 2 hours before quenched with sat Na₂CO₃.Solvents were removed under vacuum, and the residue was treated with DCM(50×3 mL). Combined organic layers were dried over Na₂SO₄, and purifiedvia chromatography (SiO₂, 40 g, 0-50%ethyl acetate/hexanes) to give adesired product (2.83 g, 97%). ¹H NMR (CHLOROFORM-d) δ: 8.48 (br. s.,1H), 7.97 (d, J=8.8 Hz, 1H), 7.02 (d, J=2.8 Hz, 1H), 6.88 (dd, J=8.9,2.9 Hz, 1H), 5.88 (s, 1H), 4.66 (s, 2H), 4.01-4.18 (m, 4H), 3.95 (t,J=6.5 Hz, 2H), 3.71-3.81 (m, 1H), 3.66 (dd, J=9.5, 4.8 Hz, 1H), 3.36 (s,3H), 2.60-2.76 (m, 1H), 1.69-1.84 (m, 2H), 1.58 (br. s., 2H), 1.39-1.51(m, 2H), 1.15-1.39 (m, 7H), 0.90 (t, J=6.7 Hz, 3H). MS (M+1): 410.30.

Example 45(R)—N-(2-Formyl-4-heptyloxyphenyI)-3-methoxymethoxy-2-methylpropanamide

(R)—N-2-(1,3-Dioxolan-2-yl)-4-heptyloxyphenyl-3-methoxymethoxy-2-methylpropanamide(2.80 g, 0.00684 mol) was dissolved in acetone (20 mL, 0.2 mol),followed by 6 M hydrogen chloride in water (5 mL) at 0° C. The reactionmixture was then gradually warmed up to 23° C. for 1 hour. NaHCO₃ (sat)was then added to give pH˜7. Acetone was removed under vacuum, and theresidue was extracted with DCM (50×3 mL). The combined organic layerswere then dried over Na₂SO₄. Removal of solvent gave the desired product(2.50 g, 100%). NMR (CHLOROFORM-d) δ: 11.00 (br. s., 1H), 9.89 (s, 1H),8.72 (d, J=9.5 Hz, 1H), 7.06-7.22 (m, 2H), 4.65 (s, 2H), 4.00 (t, J=6.7Hz, 2H), 3.74-3.87 (m, 1H), 3.66 (dd, J=9.8, 5.0 Hz, 1H), 3.35 (s, 3H),2.79 (m, 1H), 1.68-1.89 (m, 2H), 1.58 (br. s., 2H), 1.17-1.52 (m, 9H),0.91 (t, J=6.8 Hz, 3H). MS (M+1): 366.30.

Example 46 (S)-6-Heptyloxy-2-(1-methoxymethoxypropan-2-yl)quinazoline

(R)—N-(2-Formyl-4-heptyloxyphenyl)-3-methoxymethoxy-2-methylpropanamide(2.50 g, 0.00684 mol) was dissolved in methanol (200 mL, 5 mol), andcooled to −78° C., followed by ammonia (20 g, 1 mol) bubbling for 3hours. The reaction mixture was then transferred to a pre-cooled highpressure reactor, and heated at 130° C. for 15 hours (310 psi). Solventwas removed, and the crude mixture was purified via chromatography(SiO₂, 40 g, 0-60% ethyl acetate/hexanes) to give the desired product(1.78 g, 75%). ¹H NMR (CHLOROFORM-d) δ: 9.26 (s, 1H), 7.90 (d, J=9.3 Hz,1H), 7.52 (dd, J=9.2, 2.6 Hz, 1H), 7.10 (d, J=2.3 Hz, 1H), 4.55-4.68 (m,2H), 4.02-4.16 (m, 3H), 3.85 (dd, J=9.4, 6.4 Hz, 1H), 3.45-3.61 (m, 1H),3.27 (s, 3H), 1.79-1.95 (m, 2H), 1.45-1.57 (m, 2H), 1.27-1.45 (m, 9H),0.91 (t, J=6.7 Hz, 3H). MS (M+1): 347.30.

Example 47 (S)-2-(6-Heptyloxyquinazolin-2-yl)propan-1-ol

(S)-6-Heptyloxy-2-(1-methoxymethoxypropan-2-yl)quinazoline (1500 mg,0.0043 mol) was dissolved in methanol (60 mL, 1 mol), followed by 6 Mhydrogen chloride in water (20 mL) and at 0° C. The reaction mixture wasthen heated at 80° C. for 30 minutes. Saturated K₂CO₃ was then added insmall portions (pH˜8), and MeOH was removed under vacuum. The aqueouslayer was treated with DCM (50×5 mL). The combined organic layers werethen washed with water and brine, and then dried over Na₂SO₄. The crudemixture was purified via chromatography (SiO₂, 4 g, 0-100% ethylacetate/hexanes) to give 1.10 g (84%), and 189 mg of methyl ether sideproduct. ¹H NMR (CHLOROFORM-d) δ: 9.25 (s, 1H), 7.88 (d, J=9.3 Hz, 1H),7.49-7.62 (m, 1H), 7.12 (d, J=2.5 Hz, 1H), 4.18-4.29 (m, 1H), 4.04-4.15(m, 3H), 3.90-4.03 (m, 1H), 3.39 (m, 1H), 1.79-1.97 (m, 2H), 1.21-1.60(m, 11H), 0.92 (t, J=6.8 Hz, 3H). MS (M+1): 303.30.

Example 48 (S)-2-(6-Heptyloxyquinazolin-2-yl)propyl carbamate

(S)-2-(6-Heptyloxy-quinazolin-2-yl)-propan-1-ol (13, 950 mg, 0.0031 mol)was dissolved in methylene chloride (100 mL, 2 mol), followed bytrichloroacetyl isocyanate (449 μL, 0.00377 mol) dropwise at 0° C. Thereaction mixture was then gradually warmed up to 23° C., and stirred foradditional for 1 hour. Solvent was removed under vacuum, and the residuewas redissolved in methanol (100 mL, 3 mol), followed by potassiumcarbonate (5000 mg, 0.03 mol) and water (20 mL, 1 mol) at 0° C. Thereaction mixture was then warmed up to at 23° C. and stirred for 3hours. MeOH was removed under vacuum, and the aqueous layer was treatedwith DCM (10×5 mL). The combined organic layers were then washed withwater and brine, and then dried over Na₂SO₄. The crude mixture waspurified via chromatography (SiO₂, 4 g, 0-100% ethyl acetate/hexanes) togive 1.05 g, (97%). ¹H NMR (400 MHz, CHLOROFORM-d) δ: 9.24 (s, 1H), 7.89(d, J=9.3 Hz, 1H), 7.53 (dd, J=9.0, 2.8 Hz, 1H), 7.11 (d, J=2.8 Hz, 1H),4.60 (dd, J=10.5, 8.0 Hz, 1H), 4.51 (br. s., 2H), 4.45 (dd, J=10.5, 6.0Hz, 1H), 4.09 (t, J=6.5 Hz, 2H), 3.47-3.62 (m, 1H), 1.81-1.92 (m, 2H),1.40-1.65 (m, 11H), 0.82-0.96 (m, 3H). MS (ES, M+1): 346.30.

Example 49 (R)-4-(6-Heptyloxyquinazolin-2-yl)-4-methyloxazolidin-2-one

(S)-2-(6-Heptyloxyquinazolin-2-yl)propyl carbamate (14, 350 mg, 0.0010mol) was dissolved in toluene (20 mL, 0.2 mol), followed by magnesiummonoxide (93.9 mg, 0.00233 mol), iodobenzene diacetate (359 mg, 0.00111mol) and finally Rh₂(esp)₂ (35 mg, 0.000046 mol). The reaction mixturewas then heated at 40° C. for 3 days. DCM was added to the reactionmixture, and the reaction mixture was filtered. The crude mixture wasthen purified via chromatography (SiO₂, 12 g, 0-80% ethylacetate/hexanes) to give 53 mg of the desired product (15%) and 215.0 mgof 14. ¹H NMR (CHLOROFORM-d) δ: 9.27 (s, 1H), 7.90 (d, J=9.3 Hz, 1H),7.58 (dd, J=9.2, 2.6 Hz, 1H), 7.15 (d, J=2.5 Hz, 1H), 6.19 (br. s., 1H),5.18 (d, J=8.5 Hz, 1H), 4.49 (d, J=8.8 Hz, 1H), 4.11 (t, J=6.5 Hz, 2H),1.80-1.97 (m, 2H), 1.76 (s, 3H), 1.21-1.60 (m, 8H), 0.91 (t, J=6.7 Hz,3H). MS (M+1): 344.30.

Example 50 (R)-2-Amino-2-(6-heptyloxyquinazolin-2-yl)propan-1-ol

(R)-4-(6-Heptyloxy-quinazolin-2-yl)-4-methyl-oxazolidin-2-one (40.0 mg,0.000116 mol) was dissolved in ethanol (2 mL, 0.03 mol), followed by 4 Mlithium hydroxide in water (1 mL). The reaction mixture was then heatedat 80° C. for 2 hours. All solvent was removed. The solid was extractedwith DCM, and was purified via chromatography (SiO₂, 4 g, 0-100% (10% ofMeOH with 2N ammonia in DCM and DCM) to give the desired product (21 mg,57%). ¹H NMR (CHLOROFORM-d) δ: 9.25 (s, 1H), 7.87 (d, J=9.0 Hz, 1H),7.49 (dd, J=9.7, 2.4 Hz, 1H), 7.08 (d, J=2.5 Hz, 1H), 6.60 (br. s., 3H),4.26 (d, J=11.8 Hz, 1H), 4.17 (d, J=11.8 Hz, 1H), 4.06 (t, J=6.7 Hz,2H), 1.81-2.00 (m, 4H), 1.77 (s, 3H), 1.43-1.61 (m, 2H), 1.20-1.43 (m,4H), 0.84-1.01 (m, 3H). MS (ES, M+1): 318.30.

Example 51 (R)-2-Amino-2-(6-(heptyloxy)quinazolin-2-yl)propyl dihydrogenphosphate

A three step procedure similar to that used for phosphoric acidmono-{2-amino-2-[6-(4-tert-butyl-cyclohexyloxy)-quinolin-2-yl]-propyl}ester Enantiomer 1 (Examples 28-30) was used to make the title compoundfrom (R)-2-amino-2-(6-heptyloxyquinazolin-2-yl)propan-1-ol. ¹H NMR(MeOD) δ: 9.49 (s, 1H), 8.01 (d, J=9.3 Hz, 1H), 7.69 (dd, J=9.2, 2.6 Hz,1H), 7.49 (d, J=2.8 Hz, 1H), 4.63 (dd, J=10.8, 4.8 Hz, 1H), 4.47 (dd,J=10.9, 4.9 Hz, 1H), 4.18 (t, J=6.4 Hz, 2H), 1.83-1.96 (m, 2H), 1.81 (s,3H), 1.48-1.60 (m, 2H), 1.23-1.47 (m, 6H), 0.84-0.99 (m, 3H). MS (M+1):398.30.

Example 52 5-Methoxy-2-nitrobenzaldehyde

5-Hydroxy-2-nitrobenzaldehyde (25.0 g, 150 mmol) was dissolved inN,N-dimethylformamide (200 mL, 2000 mmol), followed by potassiumcarbonate (20.7 g, 150 mmol) and methyl iodide (10.2 mL, 164 mmol). Thereaction mixture was stirred for 10 hours at 23° C. Ethyl acetate (1000mL) was added, and the mixture was washed with water and brine. Organiclayer was dried over sodium sulfate. Removal of solvent gave a crudesolid product, which was treated with DCM and hexanes. The solid wascollected and washed with hexanes (27.0 g, 100%). NMR (CHLOROFORM-d) δ:10.50 (s, 1H), 8.18 (d, J=9.0 Hz, 1H), 7.34 (d, J=2.8 Hz, 1H), 7.16 (dd,J=9.2, 2.9 Hz, 1H), 3.97 (s, 3H). MS (M+1): 182.10.

Example 53 2-(5-Methoxy-2-nitrophenyl)-1,3-dioxolane

5-Methoxy-2-nitro-benzaldehyde (30.00 g, 0.1656 mol) and1,2-bis-trimethylsilanyloxy-ethane (40.6 mL, 0.166 mol) were placed in a40 mL vial, followed by Methylene chloride (1000 mL, 20 mol). Thereaction mixture was then cooled to −78° C., and trimethylsilyltrifluoromethanesulfonate (1 mL, 0.008 mol) was added dropwise under N₂.The reaction mixture was allowed to warm up to at 23° C. for 1 day. Asolution of saturated K₂CO₃ (50 mL) was added to quench the reaction,and organic layer was separated. Removal of solvent gave a oily product,which was treated with ether/hexanes to give a pure solid product (37.0g, 99%). NMR (MeOD) δ: 8.01 (d, J=9.0 Hz, 1H), 7.29 (d, J=2.8 Hz, 1H),7.05 (dd, J=8.9, 2.9 Hz, 1H), 6.49 (s, 1H), 4.85 (s, 3H), 3.91 (s, 4H).MS (M+1): 226.10.

Example 54 2-(1,3-Dioxolan-2-yl)-4-methoxyaniline

2-(5-Methoxy-2-nitrophenyl)-1,3-dioxolane (15.00 g, 0.06661 mol),platinum dioxide (900 mg, 0.004 mol) and sodium acetate trihydrate (700mg, 0.005 mol) were placed in a pressure flask, followed by ethylacetate (400 mL, 4 mol). The reaction mixture was then purged under N₂for at least 3 times, and H₂ was introduced (purged 3 times) andmaintained at 52 psi for 3 hours. The reaction mixture was thenfiltered, and solvent was removed to give a pure product (13 g, 100%).¹H NMR (CHLOROFORM-d) δ: 6.96 (d, J=2.8 Hz, 1H), 6.76 (dd, J=8.5, 3.0Hz, 1H), 6.64 (d, J=8.5 Hz, 1H), 5.84 (s, 1H), 4.00-4.18 (m, 4H), 3.89(br. s., 2H). MS (M+1): 196.10.

Example 55 (S)-tert-Butyl1-(2-(1,3-dioxolan-2-yl)-4-methoxyphenylamino)-3-hydroxy-2-methyl-1-oxopropan-2-ylcarbamate

(S)-2-tert-Butoxycarbonylamino-3-hydroxy-2-methyl-propionic acid (247.1mg, 0.001127 mol) was dissolved in DMF, followed byN,N-diisopropylethylamine (0.892 mL, 0.00512 mol), and then2-(1,3-dioxolan-2-yl)-4-methoxyaniline (200.0 mg, 0.001024 mol) andN,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl)uroniumhexafluorophosphate (389.5 mg, 0.001024 mol) at 23° C. The reactionmixture was allowed to stir at 23° C. for 10 hours. Ethyl acetate (50mL) was added, followed by water (10 mL). Organic layer was separated,and the aqueous layer was extracted with additional ethyl acetate(2×10mL). Combined organic layers were washed with water (3×10 mL),brine and dried over Na₂SO₄. The crude product was purified viachromatography (SiO₂, 12 g, 0-50% ethyl acetate/hexanes) to give thedesired product (176 mg, 43%). ¹H NMR (MeOD) δ: 7.98 (s, 1H), 7.01 (d,J=3.0 Hz, 1H), 6.93 (dd, J=8.9, 2.9 Hz, 1H), 5.76 (s, 1H), 4.13-4.26 (m,2H), 3.95-4.09 (m, 2H), 3.69-3.85 (m, 7H), 1.37-1.53 (m, 12H). MS (M+1):397.20.

Example 56 (R)-2-Amino-2-(6-methoxyquinazolin-2-yl)propan-1-ol

[(S)-1-(2-1,3-Dioxolan-2-yl-4-methoxy-phenylcarbamoyl)-2-hydroxy-1-methyl-ethyl]-carbamicacid tert-butyl ester (20 g, 50 mmol) was dissolved in MeOH (50 mL), andcooled to around −30° C. 6 M Hydrogen chloride in water (10.09 mL, 60.54mmol) was added and the reaction mixture was allowed to warm to at 23°C. over for 2 hours. All the solvents were removed under vacuum and theresidue was re-dissolved in MeOH (20mL) and cooled to −78° C. Thissolution was then added to a solution of ammonia in MeOH (80 mL, withammonia gas bubbled through for 3 h at at −78° C.). The mixture was thentransferred to a high pressure vessel, and heated at 130° C. for 6 hours(320 psi). The mixture was filtrated and the residue was purified viachromatography (SiO₂, 120 g, 0-20% MeOH/DCM) to give 3.1 g of pureproduct (30%). ¹H NMR (CHLOROFORM-d) δ: 9.28 (s, 1H), 7.90 (d, J=9.4 Hz,1H), 7.56 (dd, J=9.3, 2.8 Hz, 1H), 7.14 (d, J=2.6 Hz, 11-1), 4.07 (d,J=11.0 Hz, 1H), 3.97 (s, 3H), 3.84 (d, J=10.6 Hz, 1H), 2.67 (br. s.,3H), 1.57 (s, 3H). MS (M+1): 234.10.

Example 57 (R)-4-(6-Methoxyquinazolin-2-yl)-4-methyloxazolidin-2-one

(R)-2-Amino-2-(6-methoxy-quinazolin-2-yl)-propan-1-ol (3.10 g, 13.3mmol) was dissolved in methylene chloride (60 mL, 900 mmol), followed byN,N-diisopropylethylamine (6.94 mL, 39.9 mmol). The mixture was thencooled down to −78° C., and triphosgene (4.34 g, 14.6 mmol) was added insmall portions over for 1 hour. The reaction mixture was then warmed upto 23° C. gradually and stirred for additional for 3 hours. Quenchedwith K₂CO₃ (sat), and organic layer was separated. The aqueous layer wasextracted with DCM (2×20 mL) and combined organic layers were washedwith water, brine and dried over Na₂SO₄. The crude product was thenpurified via chromatography (SiO₂, 40 g, 0-10% MeOH/DCM) (1.78 g, 51%).¹H NMR (CHLOROFORM-d) δ: 9.29 (s, 1H), 7.90 (d, J=9.4 Hz, 1H), 7.58 (dd,J=9.1, 2.6 Hz, 1H), 7.16 (d, J=2.6 Hz, 1H), 6.29 (br. s., 1H), 5.18 (d,J=8.7 Hz, 1H), 4.49 (d, J=8.7 Hz, 1H), 3.97 (s, 3H), 1.76 (s, 3H). MS(M+1): 260.10.

Example 58 (R)-4-(6-Hydroxyquinazolin-2-yl)-4-methyloxazolidin-2-one

(R)-4-(6-Methoxy-quinazolin-2-yl)-4-methyl-oxazolidin-2-one (860.0 mg,3.317 mmol) was dissolved in methylene chloride (60 mL, 900 mmol), andcooled to −78° C. A solution of 1.0 M of boron tribromide in nethylenechloride (9.95 mL, 9.95 mmol) was then added dropwise. The reactionmixture was then warmed up to at 23° C. gradually and then heated at 50°C. for 2 hours. The reaction mixture was then question with NaHCO₃ (sat)to basic, and neutralized back to pH-7-8. DCM was removed under vacuum,the white solid was collected and washed with water and hexanes (560 mg,69%). ¹H NMR (MeOD) δ: 9.32 (s, 1H), 7.91 (d, J=9.3 Hz, 1H), 7.57 (dd,J=9.0, 2.8 Hz, 1H), 7.26 (d, J=2.5 Hz, 1H), 4.98 (d, J=8.5 Hz, 1H), 4.46(d, J=8.5 Hz, 1H), 1.78 (s, 3H). MS (M+1): 246.10.

Example 59(R)-4-(6-Hydroxy-5-iodoquinazolin-2-yl)-4-methyloxazolidin-2-one

(R)-4-(6-Hydroxy-quinazolin-2-yl)-4-methyl-oxazolidin-2-one (330.0 mg,0.001346 mol) was dissolved in methylene chloride (500 mL, 8 mol),(heterogeneous) followed by N-iodosuccinimide (302.8 mg, 0.001346 mol)at 23° C. The reaction mixture was then sonicated for 30 seconds, andall the solvent was removed under vacuum (rotavap). The residue waspurified via chromatography (SiO₂, 20 gm, 0-100% ethyl acetate/hexanes)to give 420 mg of desired product (84%). ¹H NMR (MeOD) δ: 9.50 (s, 1H),7.90 (d, J=9.0 Hz, 1H), 7.60 (d, J=9.0 Hz, 1H), 4.99 (d, J=8.8 Hz, 1H),4.48 (d, J=8.8 Hz, 1H), 1.79 (s, 3H). MS (M+1): 372.00.

Example 59A(R)-4-(6-(trans-4-tert-Butylcyclohexyloxy)-5-iodoquinazolin-2-yl)-4-methyloxazolidin-2-one

(R)-4-(6-Hydroxy-5-iodo-quinazolin-2-yl)-4-methyl-oxazolidin-2-one(350.0 mg, 0.0009431 mol), methanesulfonic acid 4-tert-butyl-cyclohexylester (884.0 mg, 0.003772 mol) and cesium carbonate (921.8 mg, 0.002829mol) were dissolved in a mixture of tert-butyl alcohol (19.9 mL, 0.208mol) and 2-butanone (6.6 mL, 0.074 mol). The reaction mixture was heatedat 80° C. for 1 hour (MW), an additional methanesulfonic acid4-tert-butyl-cyclohexyl ester (500 mg) was added, and the mixture washeated at 80° C. for an additional for 1 hour. LC/MS indicated thatreaction was completed. Removal of solvents gave a solid mixture, whichwas treated with DCM and filtered. The filtrate was then washed withwater, brine and dried over Na₂SO₄. The crude mixture was then purifiedvia chromatography (SiO₂, 24 gm, 0-100% ethyl acetate/hexanes) to give451 mg desired product (94%).¹H NMR (CHLOROFORM-d) δ: 9.55 (s, 1H), 7.94(d, J=9.3 Hz, 1H), 7.61 (d, J=9.3 Hz, 1H), 6.07 (s, 1H), 5.16 (d, J=8.8Hz, 1H), 4.50 (d, J=8.8 Hz, 1H), 4.24-4.42 (m, 1H), 2.24 (d, J=12.0 Hz,2H), 1.92 (d, J=9.5 Hz, 2H), 1.76 (s, 3H), 1.63 (d, J=11.3 Hz, 1H),1.22-1.39 (m, 2H), 1.15 (br. s., 2H), 0.81-0.98 (m, 9H). MS (M+1):510.10.

Example 60(R)-4-(6-(trans-4-tert-Butylcyclohexyloxy)-5-(trifluoromethyl)quinazolin-2-yl)-4-methyloxazolidin-2-one

(R)-4-[6-(4-tert-Butyl-cyclohexyloxy)-5-iodo-quinazolin-2-yl]-4-methyl-oxazolidin-2-one(360.0 mg, 0.0007067 mol) and copper(I)iodide (202 mg, 0.00106 mol) wereplaced in a vial, and purged with N₂. N,N-cimethylformamide (10.0 mL,0.129 mol) was added followed by hexamethylphosphoramide (0.700 mL,0.00402 mol). Methyl fluorosulphonyldifluoroacetate (0.900 mL, 0.00707mol) was added dropwise at 23° C. The reaction mixture was then heatedto 80° C. for 40 min. The reaction mixture was then cooled to at 23° C.,and filtrated. Majority of the solvent was removed under vacuum, andwater was added to the residue solution and extracted with DCM (25×3mL). Combined organic layers were dried over Na₂SO₄, and purified viachromatography (SiO₂, 12 gm, 0-50% ethyl acetate/hexanes) to give 197 mgof the desired product (62%). ¹H NMR (CHLOROFORM-d) δ: 9.75 (s, 1H),8.12 (d, J=9.4 Hz, 1H), 7.76 (d, J=9.4 Hz, 1H), 6.18 (s, 1H), 5.14 (d,J=9.1 Hz, 1H), 4.48 (d, J=8.7 Hz, 1H), 4.27-4.45 (m, 1H), 2.20 (d,J=11.3 Hz, 2H), 1.81-2.00 (m, 2H), 1.76 (s, 3H), 1.45-1.68 (m, 1H),1.03-1.36 (m, 4H), 0.78-0.94 (m, 9H). MS (M+1): 452.20.

Example 61(R)-2-Amino-2-(6-(trans-4-tert-butylcyclohexyloxy)-5-(trifluoromethyl)quinazolin-2-yl)propan-1-ol

(R)-4-[6-(4-tert-Butyl-cyclohexyloxy)-5-trifluoromethyl-quinazolin-2-yl]-4-methyl-oxazolidin-2-one(180.0 mg, 0.0003987 mol) was dissolved in ethanol (7 mL, 0.1 mol),followed by 4 M lithium hydroxide in water (3 mL, 0.01 mol). Thereaction mixture was then heated at 80° C. for 2 hours. All solvent wasremoved. The solid was extracted with DCM, and organic layers were driedover Na₂SO₄. Removal of solvent gave a pure product (152 mg, 90%). ¹HNMR (MeOD) δ: 9.73 (s, 1H), 8.24 (d, J=9.3 Hz, 1H), 8.05 (d, J=9.5 Hz,1H), 4.50-4.68 (m, 1H), 4.11 (d, J=11.0 Hz, 1H), 3.84 (d, J=11.0 Hz,1H), 2.22 (d, J=10.8 Hz, 2H), 1.92 (d, J=13.1 Hz, 2H), 1.61 (s, 3H),1.43-1.58 (m, 2H), 1.18-1.35 (m, 2H), 1.04-1.18 (m, 1H), 0.91 (s, 9H).MS (M+1): 426.30.

Example 62(R)-2-amino-2-(6-(trans-4-tert-butylcyclohexyloxy)-5-(trifluoromethyl)quinazolin-2-yl)propyldihydrogen phosphate

A three step procedure similar to that used for phosphoric acidmono-{2-amino-2-[6-(4-tert-butyl-cyclohexyloxy)-quinolin-2-yl]-propyl}ester Enantiomer 1 (Examples 28-30) was used to make the title compoundfrom(R)-2-amino-2-(6-(trans-4-tert-butylcyclohexyloxy)-5-(trifluoromethyl)quinazolin-2-yl)propan-1-ol.¹H NMR (MeOD) δ: 9.81 (s, 1H), 8.29 (d, J=9.5 Hz, 1H), 8.13 (d, J=9.5Hz, 1H), 4.55-4.70 (m, 2H), 4.44 (dd, J=10.9, 4.9 Hz, 1H), 2.23 (d,J=11.0 Hz, 2H), 1.93 (d, J=13.1 Hz, 2H), 1.81 (s, 3H), 1.43-1.65 (m,2H), 1.18-1.37 (m, 2H), 1.04-1.19 (m, 1H), 0.92 (s, 9H). MS (M+1):506.20.

Example 63(R)-2-Amino-2-(6-(trans-4-tert-butylcyclohexyloxy)quinazolin-2-yl)propan-1-ol

Coupling of t-butyl cyclohexane to the core was performed similar tothat described for(R)-4-(6-(trans-4-tert-butylcyclohexyloxy)-5-iodoquinazolin-2-yl)-4-methyloxazolidin-2-one(Example 60) utilizing(R)-4-(6-hydroxyquinazolin-2-yl)-4-methyloxazolidin-2-one as thestarting material. Deprotection was then performed similar to thatdescribed for(R)-2-amino-2-(6-(trans-4-tert-butylcyclohexyloxy)-5-(trifluoromethyl)quinazolin-2-yl)propan-1-ol(Example 61) to give the title compound. ¹H NMR (MeOD) δ: 9.37 (s, 1H),7.92 (d, J=9.3 Hz, 1H), 7.57 (dd, J=9.2, 2.6 Hz, 1H), 7.42 (d, J=2.5 Hz,1H), 4.34-4.49 (m, 1H), 4.05 (d, J=10.8 Hz, 1H), 3.79 (d, J=10.8 Hz,1H), 2.29 (d, J=10.8 Hz, 2H), 1.93 (d, J=12.8 Hz, 2H), 1.54 (s, 3H),1.38-1.51 (m, 2H), 1.21-1.35 (m, 2H), 1.08-1.19 (m, 1H), 0.93 (s, 9H).MS (M+1): 358.20.

Example 64 tert-butyl(R)-2-((R)-6-(trans-4-tert-butylcyclohexyloxy)-1,2,3,4-tetrahydronaphthalen-2-yl)-1-hydroxypropan-2-ylcarbamate

(R)-2-Amino-2-((R)-6-(trans-4-tert-butylcyclohexyloxy)-1,2,3,4-tetrahydronaphthalen-2-yl)propan-1-ol(Example 11) (30.0 mg, 0.0000834 mol) in chloroform (4 mL, 0.05 mol) andsaturated aqueous sodium bicarbonate solution (2 mL, 0.02 mol) was addeddi-tert-butyldicarbonate (27.3 mg, 0.000125 mol) and the mixture wasstirred at rt for 24 h. TLC shows complete reaction. After separation oforganic layer, the aqueous layer was extracted with CHCl₃. The organiclayer was washed with brine and dried over anhydrous Na₂SO₄. Theconcentrated residue was chromatographed with MeOH/CH₂Cl₂ (0-55%) togive tert-butyl(R)-2-((R)-6-(trans-4-tert-butylcyclohexyloxy)-1,2,3,4-tetrahydronaphthalen-2-yl)-1-hydroxypropan-2-ylcarbamate(38.4 mg, 100%).

Example 65(R)-2-((R)-6-(trans-4-tert-butylcyclohexyloxy)-1,2,3,4-tetrahydronaphthalen-2-yl)-1-(phosphonooxy-o-xylylene)propan-2-ylcarbamate

To a solution of tert-butyl(R)-2-((R)-6-(trans-4-tert-butylcyclohexyloxy)-1,2,3,4-tetrahydronaphthalen-2-yl)-1-hydroxypropan-2-ylcarbamate(38.4 mg, 0.0000835 mol) and 1H-tetrazole (17.6 mg, 0.000251 mol) intetrahydrofuran (0.88 mL, 0.011 mol) was added o-xylyleneN,N-diethylphosphoramidite (27.0 uL, 0.000125 mol) at rt. The resultingmixture was stirred at rt for 3 d, then hydrogen peroxide (190 uL,0.0018 mol) was added and the mixture was stirred at rt for 1 h. Thereaction was quenched with satd. NaS₂O₃, then extracted with EtOAc, thendried over Na₂SO₄. The residue was chromatographed with MeOH/CH₂Cl₂(0-100%) to give desired product (46.3mg, 86%). H1NMR confirms theidentity.

Example 66(R)-2-((R)-6-(trans-4-tert-butylcyclohexyloxy)-1,2,3,4-tetrahydronaphthalen-2-yl)-1-(phosphonooxy)propan-2-ylcarbamate

The above phosphate (46.3 mg, 0.0000721 mol) in methanol (2.0 mL, 0.049mol) was added 10% palladium on carbon (1:9, palladium:carbon black, 7.7mg). The mixture was stirred under hydrogen (0.4 L, 0.02 mol) for 2 h,filtered through Celite and washed with MeOH. The concentrated residuewas dissolved in methylene chloride and was chromatographed withMeOH/CH₂Cl₂ (0-50%) to give tert-butyl(R)-2-((R)-6-(trans-4-tert-butylcyclohexyloxy)-1,2,3,4-tetrahydronaphthalen-2-yl)-1-(phosphonooxy)propan-2-ylcarbamateas a white solid (38.0 mg, 97.6%). ¹H NMR was consistent with the titlecompound.

Example 67(R)-2-amino-2-((R)-6-(trans-4-tert-butylcyclohexyloxy)-1,2,3,4-tetrahydronaphthalen-2-yl)propyldihydrogen phosphate

tert-Butyl(R)-2-((R)-6-(trans-4-tert-butylcyclohexyloxy)-1,2,3,4-tetrahydronaphthalen-2-yl)-1-(phosphonooxy)propan-2-ylcarbamate(38.4 mg, 0.0000712 mol) was dissolved in acetic acid (2.2 mL, 0.039mol) and 10 M hydrogen chloride in water (0.6 mL) was added and themixture was stirred for 1 d. Lyophilizing gave(R)-2-amino-2-((R)-6-(trans-4-tert-butylcyclohexyloxy)-1,2,3,4-tetrahydronaphthalen-2-yl)propyldihydrogen phosphate as a white solid (16.4 mg, 52%). LCMS: Rf=1.57 min214 nm (440.22, [M+1]+, 100%). ¹H NMR (400 MHz ,MeOD) δ=7.03 (d, J=8.3Hz, 1H), 6.70 (d, J=10.9 Hz, 1H), 6.66 (s, 1H), 4.26 (dd, J=5.3, 11.1Hz, 1H), 4.18-4.09 (m, 1H), 4.06 (dd, J=4.2, 11.0 Hz, 1H), 2.98-2.64 (m,4H), 2.27-2.14 (m, 3H), 2.05 (d, J=10.2 Hz, 1H), 1.89 (d, J=12.3 Hz,2H), 1.62-1.42 (m, 1H), 1.32 (s, 3H), 1.32-1.29 (m, 1H), 1.28-1.02 (m,3H), 0.92 (s, 9H).

Example 68(R)-2-amino-2-((S)-6-(trans-4-tert-butylcyclohexyloxy)-1,2,3,4-tetrahydronaphthalen-2-yl)propyldihydrogen phosphate

The title compound (12.5 mg) was prepared from(R)-2-amino-2-((S)-6-(trans-4-tert-butylcyclohexyloxy)-1,2,3,4-tetrahydronaphthalen-2-yl)propan-1-ol(Example 12) (30.0 mg) following the four step procedure described inExamples 64-67. LCMS: Rf=1.61 min 214 nm (440.30, [M+1]+, 100%). ¹H NMR(400 MHz, MeOD) δ=6.97 (br. s., 1H), 6.67 (br. s., 1H), 6.62 (br. s.,1H), 4.22 (br. s., 1H), 4.10 (br. s., 1H), 4.03 (br. s., 1H), 3.17-2.49(m, 4H), 2.15 (br. s., 2H), 1.84 (br. s., 2H), 1.67-1.42 (m, 1H), 1.33(br. s., 3H), 1.29-1.04 (m, 3H), 0.88 (s, 9H).

Example 69(R)-4-((S)-6-(trans-4-tert-butylcyclohexyloxy)-5-iodo-1,2,3,4-tetrahydronaphthalen-2-yl)-4-methyloxazolidin-2-one

A mixture of(R)-4-((S)-6-(trans-4-tert-butylcyclohexyloxy)-1,2,3,4-tetrahydronaphthalen-2-yl)-4-methyloxazolidin-2-one(100 mg, 0.000259 mol), N-iodosuccinimide (65.4 mg, 0.000290 mol) andzirconium tetrachloride (9.1 mg, 0.000039 mol) in methylene chloride(2.13 mL, 0.0332 mol) was stirred at rt under Ar in a vial for 3 h. Theprecipitate was filtered off and the residue was purified with silicagel chromatography eluted is with EtOAc/hexane (0 to 40%) to give(R)-4-((S)-6-(trans-4-tert-butylcyclohexyloxy)-5-iodo-1,2,3,4-tetrahydronaphthalen-2-yl)-4-methyloxazolidin-2-oneas a solid (130 mg, 98%). ¹H NMR shows 1.6:1 mixture of 5-iodo and7-iodo isomers. LCMS: Rf=2.40min 512.39 ([M+1], 30%).

Example 70(R)-4-((S)-6-(trans-4-tert-butylcyclohexyloxy)-5-(trifluoromethyl)-1,2,3,4-tetrahydronaphthalen-2-yl)-4-methyloxazolidin-2-one

To a solution of(R)-4-((S)-6-(trans-4-tert-butylcyclohexyloxy)-5-iodo-1,2,3,4-tetrahydronaphthalen-2-yl)-4-methyloxazolidin-2-one(130 mg, 0.254 mmol), hexamethylphosphoramide (0.22 mL, 1.3 mmol) andcopper(I)iodide (73 mg, 0.38 mmol) in N,N-dimethylformamide (1 mL, 20mmol) was added methyl fluorosulphonyldifluoroacetate (0.17 mL, 1.3mmol). The mixture was heated at 80° C. overnight. After filtration, thesolvent was evaporated and the residue was purified with Si gelchromatography eluted with EtOAc/hexane (0 to 40%) to give a mixture of(R)-4-((S)-6-(trans-4-tert-butylcyclohexyloxy)-5-(trifluoromethyl)-1,2,3,4-tetrahydronaphthalen-2-yl)-4-methyloxazolidin-2-one and its 7-CF₃ isomer (1.8:1 shown by ¹HNMR) as a gel (95 mg, 82%). LCMS: Rf=2.41min, 450.28 (80%).

Example 71(R)-2-amino-2-((S)-6-(trans-4-tert-butylcyclohexyloxy)-5-(trifluoromethyl)-1,2,3,4-tetrahydronaphthalen-2-yl)propan-1-oland Example 71A(R)-2-amino-2-((S)-6-(trans-4-tert-butylcyclohexyloxy)-7-(trifluoromethyl)-1,2,3,4-tetrahydronaphthalen-2-yl)propan-1-ol

(R)-4-((S)-6-(trans-4-tert-butylcyclohexyloxy)-5-(trifluoromethyl)-1,2,3,4-tetrahydronaphthalen-2-yl)-4-methyloxazolidin-2-oneand its 7-CF₃ isomer (95.0 mg, 0.000209 mol) and lithium hydroxide (55.2mg, 0.00230 mol) in ethanol (1.3 mL, 0.023 mol) and water (0.44 mL,0.025 mol) was heated to reflux for overnight. The solvent was removedunder vacuum and the residue was partitioned between water/CH₂Cl₂. Theaqueous was extensively extracted with CH₂Cl₂ and the combined organicphase was dried over Na₂SO₄. The concentrated residue was taken up intoCH₂Cl₂ and subjected to chromatography purification with MeOH/CH₂Cl₂(10:90 to 80:20) to give the product (64.1 mg, 72%) as a mixture of(R)-2-amino-2-((S)-6-(trans-4-tert-butylcyclohexyloxy)-5-(trifluoromethyl)-1,2,3,4-tetrahydronaphthalen-2-yl)propan-1-oland its 7-CF₃ isomer (1.8:1 shown by ¹H NMR). LCMS: Rf=1.79min 428.17([M+1], 100%). The mixture was subjected to SFC separation (ChiralpakAD-H (2×15 cm) 08-9743, 20% methanol (0.1% DEA)/CO₂, 100 bar, 50 mL/min)yielded 20 mg of(R)-2-amino-2-((S)-6-(trans-4-tert-butylcyclohexyloxy)-5-(trifluoromethyl)-1,2,3,4-tetrahydronaphthalen-2-yl)propan-1-ol

(Rf=1.82 min, chemical purity>97%, ee>99%). LCMS: Rf=1.79 min 428.17([M+1], 100%). ¹H NMR (400 MHz, MeOD) δ=7.22 (br. s., 1H), 6.96 (br. s.,1H), 4.20 (br. s., 1H), 3.53 (d, J=11.0 Hz, 1H), 3.46 (d, J=11.0 Hz,1H), 3.11 (d, J=17.1 Hz, 1H), 2.81 (d, J=15.2 Hz, 2H), 2.63 (t, J=13.9Hz, 1H), 2.22-2.03 (m, 3H), 1.87 (d, J=11.0 Hz, 2H), 1.82-1.70 (m, 1H),1.51-1.07 (m, 6H), 1.05 (br. s., 3H), 0.91 (br. s., 9H).

and 21 mg of(R)-2-amino-2-((S)-6-(trans-4-tert-butylcyclohexyloxy)-7-(trifluoromethyl)-1,2,3,4-tetrahydronaphthalen-2-yl)propan-1-ol(Rf=3.53 min, chemical purity>98%, ee>99%). LCMS: Rf=1.79 min 428.16([M+1], 100%). ¹H NMR (400 MHz, CHLOROFORM-d) δ=7.27 (s, 1H), 6.72 (s,1H), 4.15 (tt, J=4.4, 10.8 Hz, 1H), 3.49 (d, J=10.4 Hz, 1H), 3.44 (d,J=10.4 Hz, 1H), 2.93-2.72 (m, 3H), 2.64-2.52 (m, 1H), 2.18 (dd, J=3.0,12.8 Hz, 2H), 1.97 (ddd, J=2.4, 5.0, 12.5 Hz, 1H), 1.86 (d, J=10.0 Hz,2H), 1.81-1.71 (m, 1H), 1.66 (br. s., 4H), 1.51-1.34 (m, 3H), 1.19-1.11(m, 1H), 1.11-1.06 (m, 3H), 0.88 (s, 9H).

Example 72 tert-butyl(R)-2-((S)-6-((trans)-4-tert-butylcyclohexyloxy)-5-(trifluoromethyl)-1,2,3,4-tetrahydronaphthalen-2-yl)-1-hydroxypropan-2-ylcarbamate

(R)-2-amino-2-((S)-6-(trans-4-tert-butylcyclohexyloxy)-5-(trifluoromethyl)-1,2,3,4-tetrahydronaphthalen-2-yl)propan-1-ol(Example 70) (30.4 mg, 0.0000711 mol) in chloroform (3 mL, 0.04 mol) andsaturated aqueous sodium bicarbonate solution (2 mL, 0.02 mol) was addeddi-tert-butyldicarbonate (23.3 mg, 0.000107 mol) and the mixture wasstirred at rt for 24 h. TLC shows complete reaction. After separation oforganic layer, the aqueous layer was extracted with CHCl₃. The organiclayer was washed with brine and dried over anhydrous Na₂SO₄. Theconcentrated residue was chromatographed with MeOH/CH₂Cl₂ (0-55%) togive tert-butyl(R)-2-((S)-6-((trans)-4-tert-butylcyclohexyloxy)-5-(trifluoromethyl)-1,2,3,4-tetrahydronaphthalen-2-yl)-1-hydroxypropan-2-ylcarbamate(37 mg, 99%).

Example 73 tert-Butyl(R)-2-((S)-6-(trans-4-tert-butylcyclohexyloxy)-5-(trifluoromethyl)-1,2,3,4-tetrahydronaphthalen-2-yl)-1-(phosphonooxy-o-xylylene)propan-2-ylcarbamate

To a solution of tert-butyl(R)-2-((S)-6-((1r,4S)-4-tert-butylcyclohexyloxy)-5-(trifluoromethyl)-1,2,3,4-tetrahydronaphthalen-2-yl)-1-hydroxypropan-2-ylcarbamate(37.5 mg, 0.0000711 mol) and 1H-tetrazole (14.9 mg, 0.000213 mol) intetrahydrofuran (0.75 mL, 0.0092 mol) was added o-xylyleneN,N-diethylphosphoramidite (23.0 uL, 0.000107 mol) at rt. The resultingmixture was stirred at rt for 1 d, then hydrogen peroxide (160 μL,0.0016 mol) was added and the mixture was stirred at rt for 1 h. Thereaction was quenched with satd. NaS₂O₃, then extracted with EtOAc, thendried over Na₂SO₄. The residue was chromatographed with MeOH—CH₂Cl₂(0-100%) to give desired phosphate (50 mg, 100%). ¹H NMR was consistentwith the title compound.

Example 74 tert-butyl(R)-2-((S)-6-(trans-4-tert-butylcyclohexyloxy)-5-(trifluoromethyl)-1,2,3,4-tetrahydronaphthalen-2-yl)-1-(phosphonooxy)propan-2-ylcarbamate

To a solution of tert-Butyl(R)-2-((S)-6-(trans-4-tert-butylcyclohexyloxy)-5-(trifluoromethyl)-1,2,3,4-tetrahydronaphthalen-2-yl)-1-(phosphonooxy-o-xylylene)propan-2-ylcarbamate(50.4 mg, 0.0000710 mol) in methanol (2.0 mL, 0.048 mol) and was added10% palladium on carbon (1:9, palladium:carbon black, 7.6 mg, 0.0000071mol). The mixture was stirred under hydrogen (0.4 L, 0.02 mol) for 2 h.Filtered through celite and was washed with MeOH. The concentratedresidue was dissolved in CH₂Cl₂ and was chromatographed with MeOH/CH₂Cl₂(0-50%) to give tert-butyl(R)-2-((S)-6-(trans-4-tert-butylcyclohexyloxy)-5-(trifluoromethyl)-1,2,3,4-tetrahydronaphthalen-2-yl)-1-(phosphonooxy)propan-2-ylcarbamateas a white solid (22.0 mg, 51%). ¹H NMR was consistent with the titlecompound.

Example 75(R)-2-amino-2-((S)-6-(trans-4-tert-butylcyclohexyloxy)-5-(trifluoromethyl)-1,2,3,4-tetrahydronaphthalen-2-yl)propyldihydrogen phosphate

tert-Butyl(R)-2-((S)-6-(trans-4-tert-butylcyclohexyloxy)-5-(trifluoromethyl)-1,2,3,4-tetrahydronaphthalen-2-yl)-1-(phosphonooxy)propan-2-ylcarbamate(22.0 mg, 0.0000362 mol) was dissolved in trifluoroacetic Acid (1.0 mL,0.013 mol) and methylene chloride (1.0 mL, 0.016 mol) was added and themixture was stirred for 1 h. Concentration and lypholyzing gave pure(R)-2-amino-2-((S)-6-(trans-4-tert-butylcyclohexyloxy)-5-(trifluoromethyl)-1,2,3,4-tetrahydronaphthalen-2-yl)propyldihydrogen phosphate as a white solid (18.0 mg, 98%). LCMS: Rf=1.74min508.41 [M+1]. ¹H NMR (400 MHz, MeOD) δ=7.25 (d, J=9.0 Hz, 1H), 7.00 (d,J=8.6 Hz, 1H), 4.23 (dd, J=4.9, 11.2 Hz, 1H), 4.18 (m, 1H), 4.03 (dd,J=4.1, 11.3 Hz, 1H), 3.22-2.82 (m, 3H), 2.77-2.64 (m, 1H), 2.15 (d,J=12.6 Hz, 2H), 2.02 (d, J=10.6 Hz, 1H), 1.87 (d, J=13.2 Hz, 2H),1.56-1.35 (m, 2H), 1.33 (s, 3H), 1.24-1.01 (m, 2H), is 0.89 (s, 9H).

Example 76 6-bromo-2-(trans-4-tert-butylcyclohexyloxy)quinoline

The mixture of 6-bromo-quinolin-2-ol (1.00 g, 0.00446 mol),cis-4-tert-butylcyclohexanol (0.837 g, 0.00536 mol), andtriphenylphosphine (1.405 g, 0.005356 mol) in toluene (9.508 mL, 0.08926mol) was heated to reflux, and diisopropyl azodicarboxylate (1.054 mL,0.005356 mol) was added dropwise and was stirred at reflux for 6 hours.The mixture was taken up into methylene chloride and subjected tochromatography with EtOAc/hexane (0:100 to 40:60) to give6-bromo-2-(trans-4-tert-butylcyclohexyloxy)quinoline as a white solid(0.581 g, 36%). LCMS: Rf=2.82 min (362.38, [M]+, 100%).

Example 77 2-Methyl-propane-2-sulfinic acid[(R)-1-[2-(4-trans-tert-butylcyclohexyloxy)-quinolin-6-yl]-2-(tert-butyl-dimethyl-silanyloxy)-1-methyl-ethyl]-amide

To a solution of 6-bromo-2-(4-trans-tert-butyl-cyclohexyloxy)-quinoline(0.273 g, 0.000755 mol) in ether (1.3 mL, 0.012 mol) at −78° C. wasadded 2.0 M of n-butyllithium in cyclohexane (0.412 mL, 0.000823 mol)and stirred for 30 min then to 0° C. for 5 min. To a solution of(S)—N-(1-(tert-butyldimethylsilyloxy)propan-2-ylidene)-2-methylpropane-2-sulfinamide(0.200 g, 0.000686 mol) in toluene (6.8 mL, 0.064 mol) at −78° C. wasadded 2.0 M trimethylaluminum in toluene(0.377 mL, 0.000755 mol). Theorganolithium solution was transferred to the above mixture by syringe.And the mixture was stirred at −41° C. for 3 h. The reaction wasquenched with Na₂SO₄ aqueous saturated solution, diluted with EtOAc, andfiltrated through Celite. washed with brine, dried. The mixture wasconcentrated and crude NMR showed desired product as 1:0.28diasteroisomer mixture. The mixture was taken up into methylene chlorideand subjected to chromatography purification with EtOAc/hexane (0:100 to100:0) to give 2-methyl-propane-2-sulfinic acid[(R)-1-[2-(4-tertbutyl-cyclohexyloxy)-quinolin-6-yl]-2-(tert-butyl-dimethyl-silanyloxy)-1-methyl-ethyl]-amide(3.6:1 mixture) as a white sticky solid (0.227 g, 58%). LCMS 2.77 min575.73 ([M+1], 100%).

Example 782-amino-2-(2-(trans-4-tert-butylcyclohexyloxy)quinolin-6-yl)propan-1-ol

2-Methyl-propane-2-sulfinic acid[(R)-1-[2-(4-trans-tert-butyl-cyclohexyloxy)-quinolin-6-yl]-2-(tert-butyl-dimethyl-silanyloxy)-1-methyl-ethyl]-amide(227.4 mg, 0.3955 mmol, the above mixture) in methanol (4.5 mL, 110mmol) was added 4.0 M hydrogen chloride in 1,4-dioxane (2.2 mL, 9.0mmol) and was stirred overnight. LC showed single peak. After removal ofsolvent, the residue was dissolved in methylene chloride, 1M aq. NH₄OHwas added, and the extracted organic layer was dried. The concentratedresidue was subjected to chromatography purification with MeOH/CH₂Cl₂(20-40%) give2-amino-2-(2-(trans-4-tert-butylcyclohexyloxy)quinolin-6-yl)propan-1-olas a foamy solid (110 mg, 78%). LCMS: Rf=1.51 min 357.42([M+1]). ¹H NMR(400 MHz, MeOD) δ=8.10 (d, J=8.8 Hz, 1H), 7.86 (s, 1H), 7.77 (d, J=1.4Hz, 1H), 6.87 (d, J=8.8 Hz, 1H), 5.14 (tt, J=4.6, 11.0 Hz, 1H), 3.74 (d,J=11.0 Hz, 1H), 3.69 (d, J=11.0 Hz, 1H), 2.29 (d, J=12.5 Hz, 2H), 1.93(d, J=11.4 Hz, 2H), 1.54 (s, 3H), 1.51-1.38 (m, 2H), 1.37-1.22 (m, 2H),1.20-1.09 (m, 1H), 0.94 (s, 9H).

Example 79(R)-2-amino-2-(2-(trans-4-tert-butylcyclohexyloxy)quinolin-6-yl)propan-1-oland Example 79A(S)-2-amino-2-(2-(trans-4-tert-butylcyclohexyloxy)quinolin-6-yl)propan-1-ol

SFC separation of the above2-amino-2-[2-(4-trans-tert-butyl-cyclohexyloxy)-quinolin-6-yl]-propan-1-ol(Example 78) (100 mg, 0.3 mmol) with Whelk-01 (R,R) (3×15cm) 08-10149 in15% MeOH(0.1% DEA)/CO2, 100 bar, 85 mL/min, 220 nm gave 62 mg of peak 1(>99% ee) and 12 mg of peak 2 (>99% ee). Peak 1 was subjected tochromatography purification with CH₂Cl₂/MeOH(20-50%) give(R)-2-amino-2-(2-(trans-4-tert-butylcyclohexyloxy)quinolin-6-yl)propan-1-olas a foamy solid (39.4 mg). LCMS 1.42 min 357.30 ([M+1], 100%). ¹H NMR(400 MHz, MeOD) δ=8.10 (d, J=8.8 Hz, 1H), 7.86 (s, 1H), 7.78 (s, 2H),6.87 (d, J=8.8 Hz, 1H), 5.20-5.09 (m, 1H), 3.75 (d, J=11.0 Hz, 1H), 3.70(d, J=11.0 Hz, 1H), 2.29 (d, J=8.0 Hz, 2H), 1.93 (d, J=14.5 Hz, 2H),1.55 (s, 3H), 1.52-1.38 (m, 2H), 1.38-1.23 (m, 2H), 1.20-1.07 (m, 1H),0.94 (s, 9H).

Peak 2 was subjected to chromatography purification with MeOH/CH₂Cl₂(20-50%) give (S)-2-amino-2-(2-(trans-4-tert-butylcyclohexyloxy)quinolin-6-yl)propan-1-ol as a gel (2.0 mg). LCMS 1.42 min 357.30([M+1], 100%). NMR (400 MHz, MeOD) δ=8.10 (d, J=8.8 Hz, 1H), 7.86 (s,1H), 7.77 (d, J=1.4 Hz, 1H), 6.87 (d, J=8.8 Hz, 1H), 5.14 (tt, J=4.6,11.0 Hz, 1H), 3.74 (d, J=11.0 Hz, 1H), 3.69 (d, J=11.0 Hz, 1H), 2.29 (d,J=12.5 Hz, 2H), 1.93 (d, J=11.4 Hz, 2H), 1.54 (s, 3H), 1.51-1.38 (m,2H), 1.37-1.22 (m, 2H), 1.20-1.09 (m, 1H), 0.94 (s, 9H).

Example 80 tert-butyl(R)-2-(2-(trans-4-tert-butylcyclohexyloxy)quinolin-6-yl)-1-hydroxypropan-2-ylcarbamate

(R)-2-Amino-2-[2-(4-tert-butyl-cyclohexyloxy)-quinolin-6-yl]-propan-1-ol(20.5 mg, 0.0000575 mol) in chloroform (3 mL, 0.03 mol) and saturatedaqueous sodium bicarbonate solution (2 mL, 0.02 mol) anddi-tert-butyldicarbonate (18.8 mg, 0.0000862 mol) was added and themixture was stirred at rt for 24 h. After separation of organic layer,the aqueous layer was extracted with CHCl₃. The organic layer was washedwith brine and dried over anhydrous Na₂SO₄. The concentrated residue waschromatographed with MeOH/CH₂Cl₂ (0-55%) to give tert-butyl(R)-2-(2-(trans-4-tert-butylcyclohexyloxy)quinolin-6-yl)-1-hydroxypropan-2-ylcarbamate(26.0 mg, 100%).

Example 81{(R)-1-[2-(4-tert-Butyl-cyclohexyloxy)-quinolin-6-yl]-1-methyl-2-phosphonooxy-o-xylylene-ethyl}-carbamicacid tert-butyl ester

To a solution of{(R)-1-[2-(4-tert-Butyl-cyclohexyloxy)-quinolin-6-yl]-2-hydroxy-1-methyl-ethyl}-carbamicacid tert-butyl ester (41.6 mg, 0.0000911 mol) and 1H-tetrazole (19.1mg, 0.000273 mol) in tetrahydrofuran (0.96 mL, 0.012 mol) was addedo-xylylene N,N-diethylphosphoramidite (29.5 uL, 0.000137 mol) at rt. Theresulting mixture was stirred at rt for 1 d, then hydrogen peroxide (200μL, 0.0020 mol) was added and the mixture was stirred at rt for 1 h. Thereaction was quenched with satd. NaS₂O₃, then extracted with EtOAc, thendried over Na₂SO₄. The residue was chromatographed with MeOH/CH₂Cl₂(0-100%) to give the title compound (38.5 mg, 98%).

Example 82(R)-2-amino-2-(2-(trans-4-tert-butylcyclohexyloxy)quinolin-6-yl)propyldihydrogen phosphate

{(R)-1-[2-(4-tert-Butyl-cyclohexyloxy)-quinolin-6-yl]-1-methyl-2-phosphonooxy-o-xylylene-ethyl}-carbamicacid tert-butyl ester (38.5 mg, 0.0000603 mol) in methanol (1.7 mL,0.041 mol) and was added 10% palladium on carbon (1:9, palladium:carbonblack, 6.4 mg, 0.0000060 mol). The mixture was stirred under hydrogen(0.3 L, 0.01 mol) for 2 h. Filtrate through Celite and washed with MeOH.The concentrated residue (28.6 mg) was moved to next step directly.(R)-1-[2-(4-tert-Butyl-cyclohexyloxy)-quinolin-6-yl]-1-methyl-2-phosphonooxy-ethyl}-carbamicacid tert-butyl ester (28.6 mg, 0.0000533 mol) was dissolved inmethylene chloride (1.0 mL, 0.016 mol) and trifluoroacetic Acid (1.0 mL,0.013 mol) was added and the mixture was stirred for 1 h. LC give asingle peak. Concentration and lyophilizing gave(R)-2-amino-2-(2-(trans-4-tert-butylcyclohexyloxy)quinolin-6-yl)propyldihydrogen phosphate as a white solid (30 mg, 100%). LCMS: Rf=1.32 min254 nm (437.20, [M+1]+, 100%). ¹H NMR (400 MHz, MeOD) δ=8.17 (d, J=10.4Hz, 1H), 7.95-7.85 (m, 2H), 7.81-7.74 (m, 1H), 6.97 (d, J=8.2 Hz, 1H),5.23-5.11 (m, 1H), 4.33 (dd, J=4.0, 11.0 Hz, 1H), 4.20 (d, J=5.5 Hz,1H), 2.29 (d, J=13.0 Hz, 2H), 1.93 (d, J=12.9 Hz, 2H), 1.56-1.20 (m,4H), 1.20-1.06 (m, 1H), 0.94 (s, 9H).

Example 83 3-acetamidophenyl acetate

3-Aminophenol (19 g 0.17 mol) was dissolved in Ac₂O (162 g, 1.59 mol,9.5 eq.), and pyridine (4.9 g, 0.062 mol, 0.36 eq.) was added. Then thereaction mixture was stirred at 80° C. over 2 h. Ice water (50 mL) wasadded to the mixture, and saturated NaHCO₃ solution was added to themixture until pH=7, then extracted (EA), washed (brine), dried (Na₂SO₄),and concentrated to give 3-acetamidophenyl acetate as gray solid (30 g,yield: 93%). ESI-MS: 194 (M+H)⁺.

Example 84 2-chloro-3-formylquinolin-7-yl acetate

A three-neck flask was charged with DMF (25 mL, 0.325 mol, 3 eq.), thenPOCl₃ (70 mL, 0.758 mol, 7 eq.) was added to the DMF at 0° C. Thesolution was stirred at 0° C. for 30 min. Then 3-acetamidophenyl acetate(20.8 g, 0.108 mol) was added to the mixture at 0° C. After 30 min themixture was heated to 65° C. and stirred for 16 h. Then, the reactionmixture was added to ice water (300 mL) and neutralized with saturatedNaHCO₃ to pH=6, extracted (EA), washed (brine), dried (Na₂SO₄), filteredand evaporated to dryness to give the crude product, which was purifiedby silica gel column chromatography (EA-PE, 3:1) to give desiredcompound as gray solid (2.67 g, yield 10%). ESI-MS: 250 (M+H)⁺. NMR (300MHz, CDCl₃) δ: 2.39 (s, 3H), 7.42-7.45 (m, 1H), 7.80-7.81 (d, 1H),7.97-8.00 (d, 1H), 8.74 (s, 1H), 10.54 (s,1H).

Example 85 7-hydroxyquinoline-3-carbaldehyde

To a solution of 2-chloro-3-formylquinolin-7-yl acetate (2.9 g, 14 mmol)and Pd(PPh₃)₄ (1.6 g, 1.4 mmol, 0.1 eq.), Et₃N (17 g, 168 mmol, 16 eq.)in DMF (100 mL) was added formic to acid (3.48 g, 75.6 mmol, 5.4 eq.)dropwise over 5 min. The mixture was warmed to 110° C. over 2 h. Then,the reaction mixture was diluted (water), extracted (EA), washed(brine), dried (Na₂SO₄), filtered and evaporated to dryness to give thecrude product, which was purified by silica gel column chromatography(PE-EA, 1:1) to give desired compound as yellow solid (1.45 g, yield60%). ESI-MS: 216 (M+H)⁺. ¹H NMR (300 MHz, CDCl₃) δ: 7.26-7.32 (m, 2H),8.04-8.07 (d, 1H), 8.78 (d, 1H), 9.13 (s, 1H), 10.13 (s, 1H).

Example 86 7-(heptyloxy)quinoline-3-carbaldehyde

To a solution of 7-hydroxyquinoline-3-carbaldehyde (1.3 g, 7.5 mmol) inDMF (30 mL) was added 1-bromo-heptane (5.37 g, 30 mmol, 4 eq.) and K₂CO₃(2.0 g, 15 mmol, 2 eq.). The mixture was warmed to 60° C. over 3 h.Then, the reaction mixture was diluted (water), extracted (EA), washed(brine), dried (Na₂SO₄), filtered and evaporated to dryness to give thecrude product, which was purified by silica gel column chromatography(PE-EA, 10:1) to give desired compound as yellow solid (610 mg, yield30%). ESI-MS: 272 (M+H)⁺. ¹H NMR (300 MHz, CDCl₃) δ: 0.90 (t, 3H), 1.252(s, 3H), 1.32-1.48 (m, 6H), 1.50-1.54 (m, 2H), 1.84-1.91 (m, 2H), 4.15(t, 2H), 7.27-7.29 (m, 1H), 7.454 (s, 1H), 7.828-7.851 (d, 1H), 8.506(s, 1H), 9.286 (s, 1H), 10.17 (s, 1H).

Example 87 1-(7-(heptyloxy)quinolin-3-yl)ethanol

To a solution of 7-(heptyloxy)quinoline-3-carbaldehyde (1.6 g, 5.9 mmol)in THF (30 mL) at 0° C. was added CH₃MgI (2 g, 12 mmol, 2 eq.) dropwiseover 10 min. The mixture was warmed to rt over 8 h. Then, the reactionmixture was quenched (water), extracted (EA), washed (brine), dried(Na₂SO₄), filtered and evaporated to dryness to give the crude product,which was purified by silica gel column chromatography (PE-EA, 1:5) togive desired compound as yellow oil (1.35 g, yield 80%). ESI-MS: 288(M+H)⁺. ¹H NMR (300 MHz, CDCl₃) δ: 0.90 (t, 3H), 1.32-1.41 (m, 6H),1.43-1.49 (m, 2H), 1.56-1.58 (d, 2H), 1.78-1.85 (m, 2H), 4.02 (t, 2H),5.02-5.07 (q, 1H), 7.14-7.17 (m, 1H), 7.36 (s, 1H), 7.60-7.63 (d, 1H),8.01 (s, 1H), 8.72 (s, 1H).

Example 88 1-(7-(heptyloxy)quinolin-3-yl)ethanone

To a solution of oxalyl chloride (987 mg, 7.8 mmol, 1.5 eq.) in dryCH₂Cl₂ (40 mL) was added slowly DMSO (1.6 g, 20.8 mmol, 4 eq.) at −78°C. under N₂ After 30 min, alcohol 1-(7-(heptyloxy)quinolin-3-yl)ethanol(1.5 g, 5.2 mmol) was added dropwise at −78° C. The mixture was stirredfor 2 h at −78° C., and then Et₃N (3.2 g, 31 mmol, 6 eq.) was added at−78° C. After 20 min, the mixture was warmed to room temperature. Thenthe reaction mixture was added to water (30 mL), extracted (DCM), washed(brine), dried (Na₂SO₄), filtered and evaporated to dryness to give thecrude product, which was purified by silica gel column chromatography(PE-EA, 10:1) to give desired compound as yellow solid (1.08 g, yield73%). ESI-MS: 286 (M+H)⁺. ¹H NMR (300 MHz, CDCl₃) δ: 0.90 (t, 3H),1.32-1.54 (m, 10H), 1.84-1.89 (m, 3H), 2.71 (s, 3H), 4.14 (t, 2H), 7.25(m, 1H), 7.43-7.44 (m, 1H), 7.79-7.82 (d, 1H), 8.61 (d, 1H), 9.12 (s,1H).

Example 895-(7-(heptyloxy)quinolin-3-yl)-5-methylimidazolidine-2,4-dione

A mixture of 1-(7-(heptyloxy)quinolin-3-yl)ethanone (730 mg, 2.56 mmol),EtOH (2 mL), H₂O (3 mL), (NH₄)₂ CO₃ (1.47 g, 15.36 mmol, 6 eq.), andNaCN (251 mg, 5.12 mmol, 2 eq.) was stirred for 16 h at 60° C. Then, tothe reaction mixture was added water (20 mL), extracted (EA), washed(brine), dried (Na₂SO₄), filtered and evaporated to dryness to give thecrude product, which was purified by silica gel column chromatography(PE-EA, 1:2) to give desired compound as yellow solid (480 mg, yield53%). ESI-MS: 356 (M+H)⁺. ¹H NMR (300 MHz, CDCl₃) δ: 0.81-0.83 (t, 3H),1.25-1.32 (m, 7H), 1.42-1.46 (m, 2H), 1.75-1.82 (m, 2H), 1.91 (s, 3H),3.99-4.03 (t, 2H), 7.14-7.17 (m, 1H), 7.36 (s, 1H), 7.60-7.64 (m, 2H),8.21 (s, 1H), 9.03 (s, 1H).

Example 90 2-amino-2-(7-(heptyloxy)quinolin-3-yl)propanoic acid

A mixture of5-(7-(heptyloxy)quinolin-3-yl)-5-methylimidazolidine-2,4-dione (1 g, 2.8mmol), EtOH (2 mL), H₂O (4 mL), and NaOH (2.24 g, 56 mmol, 20 eq.) wasstirred for 4 day at 110° C. Then, to the reaction mixture was added HCl(40%) to adjust to pH=5 and then the desired compound was generated asyellow solid (740 mg, yield 80%), without further purification for nextstep. ESI-MS: 331 (M+H)⁺. ¹H NMR (300 MHz, CD₃OD) δ: 0.81-0.83 (t, 3H),1.06-1.09 (t, 3H), 1.18-1.33 (m, 8H), 1.41-1.45 (m, 2H), 1.75-1.78 (m,2H), 1.96 (s, 3H), 3.48-3.53 (m, 2H), 4.04 (m, 2H), 7.18-7.24 (m, 2H),7.79-7.81 (d, 1H), 8.39 (s, 1H), 8.89 (s, 1H).

Example 91 2-amino-2-(7-(heptyloxy)quinolin-3-yl)propan-1-ol

To a solution of crude 2-amino-2-(7-(heptyloxy)quinolin-3-yl)propanoicacid (600 mg, 1.8 mmol) in dry THF (30 mL) was added LAH (138 mg, 3.6mmol, 2 eq.) at 0° C. under N₂. The mixture was warmed to roomtemperature for 3 h. Then to the reaction mixture was added water (1mL), diluted (EA), filtered and dried (Na₂SO₄), evaporated to dryness togive the crude product, which was purified by prep-HPLC to give desiredcompound as white solid (200 mg, yield 35%). ESI-MS: 317 (M+H)⁺. ¹H NMR(300 MHz, CDCl₃) δ: 0.90 (t, 3H), 1.32-1.42 (m, 6H), 1.48-1.52 (m, 2H),1.53 (s, 3H), 1.82-1.89 (m, 2H), 2.34 (brs, 3H), 3.66-3.69 (d, 1H),3.81-3.84 (d, 1H), 4.08 (t, 2H), 7.11-7.14 (m, 1H), 7.26 (s, 1H),7.57-7.59 (d, 1H), 8.10 (s, 1H), 8.83 (s, 1H). HPLC: 214nm: 98.948%,254nm: 99.526%.

Example 92 (E)-1-nitro-4-(oct-1-enyl)benzene

1-bromo-4-nitrobenzene (2.01 g, 10 mmol), 1-ocetene (7.8 mL, 50 mmol),and Pd(OAc)₂ (50 mg, 0.2 mmol) were stirred for 10 min under nitrogenatmosphere. Then tri-o-tolylphosphine (24 mg, 0.8 mmol) was added to themixture and stirred for 10 min. Et₃N (5.5 mL, 40 mmol) was then added tothe mixture. The mixture was heated to 110° C. for 18 h. Water was addedto the mixture and extracted with EtOAc. The organic layer wasconcentrated and purified by silica gel chromatography using PE aseluent to give compound the title compound (2.121 g, 90%). ¹H NMR (300MHz, CDCl₃) δ 8.097 (d, 2H), 7.235 (d, 2H), 6.442 (s, 1H), 5.976-5.377(m, 1H), 2.231-2.052 (m, 2H), 1.710-1.246(m, 8H), 0.850 (t, 3H).

Example 93 4-octylaniline

A MeOH (20 mL) solution of compound (E)-1-nitro-4-(oct-1-enyl)benzene(2.121 g, 9.0 mmol) and Pd—C (200 mg, 0.9 mmol) was stirred at r.t. for2 h under hydrogen atmosphere. Then the mixture was filtered and thefiltrate was evaporated to give product the title compound (1.586 g,85%) as a pink liquid. ¹H NMR (300 MHz, CDCl₃) δ 6.962 (d, 2H), 6.636(d, 2H), 3.487 (b, 2H), 2.484 (t, 2H), 1.581-1.467 (m, 2H), 1.291-1.166(m, 10H), 0.874 (t, 3H).

Example 94 6-octylbenzo[d]thiazol-2-amine

4-octylaniline (500 mg, 2.5 mmol), AcOH (4 mL), and KSCN (970 mg, 10mmol) were stirred at r.t. for 10 min, and then a solution of bromine(0.13 mL, 2.5 mmol) in AcOH (2 mL) was added to the mixture over 20 min.The reaction mixture was stirred at r.t. for 8 h, then poured into coldwater and extracted with EtOAc. The organic layer was purified by silicagel chromatography using PE/EA (3/1) to give compound the title compound(460 mg, 70%). ¹H NMR (300 MHz, CDCl₃) δ7.446 (s, 1H), 7.320 (s, 1H),7.230 (d, 1H), 2.577 (t, 2H), 1.567-1.506 (m, 2H), 1.266-1.167 (m, 10H),0.847 (t, 3H).

Example 95 2-bromo-6-octylbenzo[d]thiazole

To a solution of p-TsOH.H₂O (108 mg, 0.57 mmol) in MeCN (2 mL) was addedcompound 6-octylbenzo[d]thiazol-2-amine (50 mg, 0.19 mmol). Theresulting suspension was cooled to 10-15° C. and to this was added,gradually, a solution of NaNO₂ (26 mg, 0.38 mmol) and KBr (56 mg, 0.48mmol) in H₂O (0.5 mL). The mixture was then stirred at r.t. for 1 h.Then the mixture was extracted with EtOAc and purified by silica gelchromatography using PE to give product the title compound (50 mg, 82%).¹H NMR (300 MHz, CDCl₃) δ7.880 (t, 1H), 7.585 (s, 1H), 7.286 (t, 1H),2.711 (t, 2H), 1.686-1.571 (m, 2H), 1.309-1.186 (m, 10H), 0.874 (t, 3H).

Example 96 1-(tert-butyldimethylsilyloxy)propan-2-one

To a DCM (5 mL) solution of 1-hydroxypropan-2-one (500 mg, 6.8 mmol),DMAP (41 mg, 0.34 mmol) and Et₃N (1.2 mL, 8.16 mmol) was added TBSCl(1.133 g, 7.5 mmol) at 0° C. under nitrogen atmosphere. The mixture wasstirred at r.t. for 2 h. Then the mixture was added water and extractedwith EtOAc. The organic layer was purified by silica gel chromatographyusing PE/EA (10/1) to give the title compound (950 mg, 75%). EDI-MS(M+1): 189. ¹H NMR (300 MHz, CDCl₃) δ 4.057 (s, 2H), 2.080 (s, 3H),0.832 (s, 9H), 0.001 (s, 6H).

Example 97(E)-N-(1-(tert-butyldimethylsilyloxy)propan-2-ylidene)-2-methylpropane-2-sulfinamide

1-(tert-butyldimethylsilyloxy)propan-2-one (188 mg, 1.0 mmol) andTi(OEt)₄ (0.52 mL, 2.5 mmol) were dissolved in THF (5 mL). Then2-methylpropane -2-sulfinamide (121 mg, 1.0 mmol) was added to themixture under nitrogen atmosphere. The mixture was heated to 70° C. for2 h. The mixture was poured into brine and filtered through a plug ofCelite. The organic layer was concentrated and purified by silica gelchromatography using PE/EA (5/1) to give product the title compound (190mg, 65%). EDI-MS (M+1): 292. ¹H NMR (300 MHz, CDCl₃) δ 4.152 (s, 2H),2.248 (s, 3H), 1.156 (s, 9H), 0.839 (s, 9H), 0.001 (s, 6H).

Example 98N-(1-(tert-butyldimethylsilyloxy)-2-(6-octylbenzo[d]thiazol-2-yl)propan-2-yl)-2-methylpropane-2-sulfmamide

A toluene (2 mL) solution of 2-bromo-6-octylbenzo[d]thiazole (167 mg,0.515 mmol) was cooled down to −78° C., then 2.5 N n-BuLi (0.2 mL, 0.515mmol) was added to the mixture drop wise, and the mixture was stirred at−78° C. for 30 min. The toluene (2 mL) solution of(E)-N-(1-(tert-butyldimethylsilyloxy)propan-2-ylidene)-2-methylpropane-2-sulfinamide(100 mg, 0.344 mmol) was cooled down to −78° C., then 2.0 N AlMe₃ (0.26mL, 0.515 mmol) was added to the mixture drop wise, and the mixture wasstirred at −78° C. for 30 min. The mixture of(E)-N-(1-(tert-butyldimethylsilyloxy)propan-2-ylidene)-2-methylpropane-2-sulfinamidewas added to the mixture of the lithiated2-bromo-6-octylbenzo[d]thiazole and stirred at −78° C. for 1 h. Thensaturated NH₄Cl solution was added to the mixture to quench thereaction. The mixture was extracted with EtOAc and purified by silicagel chromatography using PE/EA (5/1) to give is the title compound. (90mg, 48%). EDI-MS (M+1): 539. ¹H NMR (300 MHz, CDCl₃) δ 7.858 (d, 1H),7.626 (s, 1H), 7.252 (s, 1H), 4.031 (d, 2H), 3.850 (d, 1H), 3.470 (s,1H), 2.699 (t, 2H), 1.797 (s, 3H), 1.623 (t, 2H), 1.295-1.261 (m, 21H),0.850 (s, 9H), 0.012 (s, 6H).

Example 99 2-amino-2-(6-octylbenzo[d]thiazol-2-yl)propan-1-ol

A methanol (5 mL) solution ofN-(1-(tert-butyldimethylsilyloxy)-2-(6-octylbenzo[d]thiazol-2-yl)propan-2-yl)-2-methylpropane-2-sulfinamide(90 mg) and 4N HCl (0.325 mL, 1.3 mmol) was stirred at r.t. for 24 h.Then the mixture was extracted with EtOAc and purified by silica gelchromatography using PE/EA (1/2) to give product the title compound (11mg, 26%). EDI-MS (M+1): 321. ¹H NMR (300 MHz, CDCl₃) δ 7.840 (d, 1H),7.699 (s, 1H), 7.276 (d, 1H), 4.031 (d, 1H), 3.722 (d, 1H), 2.730 (t,2H), 2.632 (b, 3H), 1.666 (t, 2H), 1.564 (s, 3H), 1.330-1.283 (m, 1011),0.866 (t, 3H). HPLC: 96.9% (214 nm); 98.0% (254 nm).

Example 100 6-bromo-2-(heptyloxy)quinoline

To a mixture of 6-bromoquinolin-2-ol (352 mg1.57 mmol) and silvercarbonate (863 mg, 3.14 mmol, 2.0 eq.) in toluene (20 mL) was added1-iodoheptane (355 mg, 1.57 mmol, 1.0 eq.) and stirred for 3 h at 110°C. The reaction mixture was evaporated under reduced pressure to removemost of solvent, then filtered, and washed with ethyl acetate. Thefiltrate was concentrated to give crude the title compound. The crudeproduct was purified by flash chromatography using PE/EA (20/1) aseluent to give product as a red oil (300 mg, 60%). ESI-MS: 323.8 (M+H)⁺.¹H NMR (400 MHz, DMSO-d₆) δ: 7.84-7.83 (m, 2H), 7.70-7.68 (m, 2H), 6.89(d, 1H), 4.43 (t, 2H), 1.81 (q, 2H), 1.51-1.28 (m, 8H), 0.90 (t, 3H).

Example 101 2-(heptyloxy)-6-(1-nitroethyl)quinoline

6-bromo-2-(heptyloxy)quinoline (2.014 g, 6.27 mmol) and Pd₂(dba)₃ (172mg, 0.188 mmol, 0.03 eq.) were dissolved in DME (5 mL) and stirred undernitrogen atmosphere for 10 min.2-di-tert-butylphosphino-2′-methylbiphenyl (117 mg, 0.36 mmol, 0.06 eq.)was added and the mixture was stirred at r.t. for 10 min. Thennitroethane (0.9 mL, 12.54 mmol, 2.0 eq.) and cesium carbonate (2.5 g,7.5 mmol, 1.2 eq.) were added. The mixture was stirred at 50° C. undernitrogen atmosphere for 15 h. The mixture was extracted with ethylacetate and the organic layer was purified by silica gel columnchromatography using petroleum/ethyl acetate (5/1) as eluent to giveproduct as a slight red oil (1.5 g, 78%). ESI-MS: 317.1 (M+H)⁺. ¹H NMR(400 MHz, CDCl₃) δ 7.98 (d, 1H), 7.86 (d, 1H), 7.79 (s, 1H), 7.69 (q,1H), 6.93 (d, 1H), 5.77-5.74 (m, 1H), 4.46 (t, 2H), 1.98 (d, 3H),1.84-1.80 (m, 2H), 1.48-1.29 (m, 8H), 0.89 (t, 3H).

Example 102 2-(2-(heptyloxy)quinolin-6-yl)-2-nitropropan-1-ol

2-(heptyloxy)-6-(1-nitroethyl)quinoline (1.823 g, 5.77 mol) andparaformaldehyde (346 mg, 11.54 mmol, 2.0 eq.) were dissolved in THF (10mL). CH₃ONa (115 mg, 2.13 mmol, 0.37 eq.) in methanol (2 mL) was addedto the solution. The mixture was stirred at 80° C. under nitrogenatmosphere for 15 h. The mixture was extracted with ethyl acetate andthe organic layer was concentrated and purified by silica gel columnchromatography using petroleum/ethyl acetate (5/1) as eluent to giveproduct as a slight red oil (823 mg, 43%). ESI-MS: 347.1 (M+H)⁺. ¹H NMR(400 MHz, CDCl₃) δ 7.97 (d, 1H), 7.88 (d, 1H), 7.69 (d, 1H), 7.56 (q,1H), 6.93 (d, 1H), 4.62 (d, 1H), 4.47 (t, 2H), 3.90 (d, 1H), 2.15 (s,3H), 1.84-1.80 (m, 2H), 1.49-1.24 (m, 8H), 0.89 (t, 3H).

Example 103 2-amino-2-(2-(heptyloxy)quinolin-6-yl)propan-1-ol

2-(2-(heptyloxy)quinolin-6-yl)-2-nitropropan-1-ol (200 mg, 0578 mmol)was dissolved in acetic acid (5 mL). zinc (376 mg, 5.78 mmol, 10.0 eq.)was added to the mixture at 0° C. and the mixture was stirred at 20° C.for 15 h. Saturated sodium carbonate solution was added to the mixtureuntil pH=8. Then the mixture was extracted with EtOAc and the organiclayer was concentrated and purified by silica gel column chromatographyusing dichloromethane/methanol (10/1) as eluent to give product (50 mg,27%) as a slight yellow solid. ESI-MS: 317.2 (M+H)⁺. HPLC: 95.99%. ¹HNMR (400 MHz, CD₃OD) δ 8.09 (d, 1H), 7.81-7.79 (m, 2H), 7.68 (q, 1H),6.92 (d, 1H), 4.38 (t, 2H), 3.84 (d, 1H), 3.74 (d, 1H), 1.86-1.75 (m,2H), 1.70 (s, 3H), 1.43-1.25 (m, 8H), 0.85 (t, 3H).

Example 104 6-(Benzyloxy)-3,4-dihydronaphthalen-1(2H)-one

The mixture of 6-hydroxy-3,4-dihydro-2H-naphthalen-1-one (30.74 g,0.1895 mol), benzyl bromide (27.0 mL, 0.227 mol) and potassium carbonate(39.3 g, 0.284 mol) in acetone (250 mL, 3.4 mol) was heated to refluxunder an atmosphere of nitrogen for 3 hours. The mixture was cooled to10° C. with an ice bath, filtered and washed with small amount of toacetone. The filtrate was concentrated in rotavap. The resultingcrystals were collected by filtration and washed with EtOAc-hexane thenhexane to give a pale yellow solid product as the first crop. The motherliquid was concentrated in rotavapor and the residue was purified bychromatograph with EtOAc:hexane (0:100 to 20:80) to give the second cropof solid product (38.07 g, 80%). ¹H NMR (CHLOROFORM-d) δ: 8.02 (d, J=8.6Hz, 1H), 7.32-7.51 (m, 5H), 6.91 (dd, J=8.8, 2.3 Hz, 1H), 6.80 (s, 1H),5.13 (s, 2H), 2.93 (t, J=6.1 Hz, 2H), 2.56-2.68 (m, 2H), 2.12 (quin,J=6.3 Hz, 2H). MS (M+1): 253.0.

Example 105 6-(Benzyloxy)-2-bromo-3,4-dihydronaphthalen-1(2H)-one

Copper(II)bromide (43.1 g, 0.193 mol) was heated to reflux in ethylacetate (150 mL, 1.5 mol). 6-Benzyloxy-3,4-dihydro-2H-naphthalen-1-one(24.35 g, 0.09651 mol) in chloroform (150 mL, 1.9 mol) was slowly addedand the mixture was refluxed overnight. The mixture was cooled to r.t.,filtered. The filtrate was decolorized with activated carbon, andfiltered throgh Celite. The filtrate was concentrated and dried in highvacuum pump to give an oily product (32.9g, 97%). ¹H NMR (CHLOROFORM-d)δ: 8.08 (d, J=9.1 Hz, 1H), 7.30-7.50 (m, 5H), 6.96 (dd, J=8.8, 2.3 Hz,1H), 6.82 (s, 1H), 5.14 (s, 2H), 4.70 (t, J=4.0 Hz, 1H), 3.29 (dt,J=11.0, 5.4 Hz, 1H), 2.87 (dt, J=16.9, 4.2 Hz, 1H), 2.36-2.59 (m, 2H).MS (M+1): 332.0

Example 106 7-(Benzyloxy)-3-bromo-1,2-dihydronaphthalene

6-Benzyloxy-2-bromo-3,4-dihydro-2H-naphthalen-1-one (32.9 g, 0.0964 mol)was dissolved in ether (200 mL, 2 mol) and methanol (30.0 mL, 0.740mol), cooled with ice bath to internal temperature of 3° C. Sodiumtetrahydroborate (1.82 g, 0.0482 mol) was added portionwise over 15 minat 3° C.-8° C. (internal). The mixture was stirred at 2-5° C. for 1hour. Acetic acid (2.74 mL, 0.0482 mol) was added and the mixture wasquenched with water, extracted with EtOAc (100 mL), washed with water(2×), brine, dried over anhydrous sodium sulfate, filtered, andconcentrated to give a crude intermediate,6-(benzyloxy)-2-bromo-1,2,3,4-tetrahydronaphthalen-1-ol (29.5 g).

The intermediate,6-(benzyloxy)-2-bromo-1,2,3,4-tetrahydronaphthalen-1-ol, was heated toreflux with p-toluenesulfonic acid monohydrate (0.916 g, 0.00482 mol) intoluene (300 mL, 3 mol) with a Dean-Stark tube for 1 hour, cooled tor.t., washed with saturated NaHCO₃, water, brine, dried over anhydrousNa₂SO₄, filtered, evaporated in rotavapor to give a crude oil.Chromatograph with EtOAc:hexane (0:100 to 10:90) gave a solid product(17.29g, 57%). ¹H NMR (CHLOROFORM-d) δ: 7.30-7.49 (m, 5H), 6.85-6.96 (m,1H), 6.76 (m, 3H), 5.06 (s, 2H), 2.87-3.00 (m, 2H), 2.67-2.83 (m, 2H).MS (M+1): 316.0.

Example 1072-(Benzylamino)-2-(6-(benzyloxy)-3,4-dihydronaphthalen-2-yl)propane-1,3-diol

7-Benzyloxy-3-bromo-1,2-dihydro-naphthalene (21.75 g, 0.06900 mol) wasazeotroped with toluene (100 mL, 0.9 mol) and dried on the high vacuumfor 30 min. This was dissolved in tetrahydrofuran (200 mL, 2 mol),cooled with a dry ice -isopropanol bath under an atmosphere of nitrogen.1.70 M tert-Butyllithium in pentane (89.3 mL, 0.152 mol) was addedslowly, stirred for 10 min. Triisopropyl borate (79.1 mL, 0.345 mol) wasadded and the mixture was stirred for 1 hour with cooling then broughtto r.t. The reaction was quenched with ice-water. The organic layer wasseparated and the aqueous layer was extracted with EtOAc (3×50 mL). Thecombined organic layers were washed with water, brine, dried overanhydrous Na₂SO₄, filtered, concentrated and dried on the high vacuum togive a crude intermediate,6-(benzyloxy)-3,4-dihydronaphthalen-2-ylboronic acid.

The intermediate, 6-(benzyloxy)-3,4-dihydronaphthalen-2-ylboronic acid,from previous step was dissolved in ethanol (400 mL, 7 mol).1,3-Dihydroxy-2-propanone (6.22 g, 0.0690 mol) was added followed bybenzylamine (7.54 mL, 0.0690 mol). The solution turned into a yellowsuspension in 1 min. The mixture was stirred at r.t. for 2 hours underan atmosphere of nitrogen. The mixture was filtered. The filtrate wasconcentrated and the residue was dissolved in EtOAc/MeOH (300 mL/30mL),washed with saturated NaHCO₃/water (50 mL/50 mL) and the aqueous layerwas extracted with EtOAc (2×50 mL). The combined organic layers werewashed with water (50 mL), brine, dried with Na₂SO₄, filtered,concentrated and dried in high vacuum to give a red-brown solid productas the first crop. The collected solid was dissolved in DCM-MeOH (50ml/50 mL) and stirred at r.t. overnight under an atmosphere of nitrogen.The solution was concentrated. The residue was treated as the first cropwith EtOAc-MeOH-aqueous NaHCO₃ to afford the second crop of product(4.40 g, 15%). ¹H NMR (CHLOROFORM-d) δ: 6.98-7.66 (m, 13H), 6.77 (br.s., 1H), 5.05 (br. s., 2H), 3.75-4.21 (m, 6H), 2.78 (m, 2H), 1.88-2.54(m, 2H). MS (M+1): 416.0.

Example 1082-Amino-2-(6-hydroxy-1,2,3,4-tetrahydronaphthalen-2-yl)propane-1,3-diol

A 1 L pressure vessel was charged 10% Pd—C, dry Enge1 hard (10:90,palladium:carbon black, 0.966 g, 0.000908 mol) under an atmosphere ofnitrogen. The solution of2-benzylamino-2-(6-benzyloxy-3,4-dihydro-naphthalen-2-yl)-propane-1,3-diol(23.75 g, 0.05716 mol) in ethanol (300 mL, 5 mol) and ethyl acetate (300mL, 3 mol) was added under an atmosphere of nitrogen. The mixture wasconnected to a H₂ line, flashed with 50 psi of H₂ three times, thenstirred under 50 psi of H₂ for 6 hours. The mixture was continuouslystirred under 50 psi of H₂ overnight. Acetic acid (18.0 mL, 0.316 mol)was added and the mixture was stirred under 50 psi of H₂ for 30 hours.10% Pd—C, dry Engelhard (10:90, palladium:carbon black, 1.01 g, 0.000949mol) suspended in 10 mL of EtOAc was added and the mixture wascontinuously stirred under 50 psi of H₂ for 45 hours. The mixture wasfiltered through Celite, concentrated to 54 g. The material wasdissolved in ethanol (300 mL, 5 mol) and acetic acid (10.0 mL, 0.176mol). The resulted solution was added to a pressure hydrogenation flaskcharged with 10% Pd—C, dry Engelhard (10:90, palladium:carbon black,2.10 g, 0.00197 mol) under an atmosphere of Nitrogen. The mixture wasflashed with 50 psi of H₂ three times then stirred under 50 psi of H₂for 3 days. Acetic acid (10.0 mL, 0.176 mol) was added and the mixturewas continuously stirred under 50 psi of H₂ for 24 more hours. Thereaction mixture was released from the H₂ pressure, filtered throughCelite, concentrated and dried in high vacuum to give a crude product.The material was used for next step without further purification. ¹H NMR(MeOD) δ: 6.84-6.99 (m, 1H), 6.43-6.61 (m, 2H), 3.62-3.88 (m, 4H),2.55-2.94 (m, 4H), 2.17-2.35 (m, 1H), 1.91-2.11 (m, 1H), 1.68-1.82 (m,1H). MS (M+1): 238.0.

Example 109 tert-Butyl1,3-dihydroxy-2-(6-hydroxy-1,2,3,4-tetrahydronaphthalen-2-yl)propan-2-ylcarbamate

2-Amino-2-(6-hydroxy-1,2,3,4-tetrahydro-naphthalen-2-yl)-propane-1,3-diol(27.2 g, 0.0573 mol) was dissolved in tetrahydrofuran (180 mL, 2.2 mol)and water (250 mL, 14 mol). To the solution was added portionwise sodiumbicarbonate (48.1 g, 0.573 mol) (pH ˜8). The solution was cooled with anice bath. Di-tert-butyldicarbonate (25.0 g, 0.115 mol) was added and themixture was brought to r.t. overnight. Di-tert-butyldicarbonate (5.00 g,0.0229 mol) was added and the mixture was stirred for 24 hours. Themixture was extracted with EtOAc (2×100 mL), brine (2×), dried overanhydrous sodium sulfate, filtered, concentrated in rotavapor to give adark brown oily residue. Et₂O (150 mL) was added. The resulted solid wascollected by filtration and washed with Et₂O, dried in air. The filtratewas concentrated in rotavapor and subject to chromatography purificationwith DCM:MeOH (100:0 to 96:4) to give a solid product (1.03 g, 5%). ¹HNMR (MeOD) δ: 6.76-6.95 (m, 1H), 6.40-6.62 (m, 2H), 3.64-3.92 (m, 4H),2.53-2.89 (m, 4H), 2.16-2.40 (m, 1H), 1.84-2.11 (m, 2H), 1.45 (s, 9H).MS (M+1): 338.0.

Example 110 tert-Butyl5-(6-hydroxy-1,2,3,4-tetrahydronaphthalen-2-yl)-2,2-dimethyl-1,3-dioxan-5-ylcarbamate

tert-Butyl1,3-dihydroxy-2-(6-hydroxy-1,2,3,4-tetrahydronaphthalen-2-yl)propan-2-ylcarbamate(0.400 g, 0.00118 mol) was treated with 2,2-dimethoxypropane (20.0 mL,0.163 mol) and p-toluenesulfonic acid monohydrate (0.0226 g, 0.000118mol) in ethyl acetate (20.0 mL, 0.205 molar r.t. for 24 hours. Thesolvent was removed. 2,2-dimethoxypropane (20.0 mL, 0.163 mol) wasadded, followed by boron trifluoride etherate (0.150 mL, 0.00118 mol).The mixture was stirred at r.t. overnight. The mixture was quenched withsaturated NaHCO₃ aqueous solution, extracted with EtOAc, washed withbrine, dried over Na₂SO₄, filtered, and concentrated to give a crude.Chromatograph with DCM:MeOH (100:0 to 93:7) gave a solid product (560mg, 100%). ¹H NMR (CHLOROFORM-d) δ: 6.89-6.99 (m, 1H), 6.49-6.65 (m,2H), 6.46-6.47 (s, br, 1H), 3.99 (m, 4H), 3.13-3.30 (m, 4H), 2.69-2.85(m, 2H), 2.10-2.39 (m, 1H), 1.56 (s, 6H), 1.37-1.50 (s, br, 9H). MS(M+Na+): 400.0.

Example 1116-(5-(tert-Butoxycarbonylamino)-2,2-dimethyl-1,3-dioxan-5-yl)-5,6,7,8-tetrahydronaphthalen-2-yltrifluoromethanesulfonate

[5-(6-Hydroxy-1,2,3,4-tetrahydro-naphthalen-2-yl)-2,2-dimethyl-1,3-dioxinan-5-yl]-carbamicacid tert-butyl ester (0.560 g, 0.00119 mol) was dissolved in methylenechloride (20 mL, 0.3 mol). N,N-Diisopropylethylamine (0.620 mL, 0.00356mol) was added followed by N-phenylbis(trifluoromethanesulphonimide)(0.594 g, 0.00166 mol). The solution was stirred at r.t. for 22 hours.The mixture was concentrated to ˜5 mL, chromatographed with EtOA:hexane(0:100 to 40:60) and a solid product was obtained (313 mg, 52%). ¹H NMR(CHLOROFORM-d) δ: 7.11 (m, 1H), 6.95 (m, 2H), 4.81 (s, br, 1H), 3.95 (m,4H), 3.20 (m, 1H), 2.86 (m, 3H), 2.67 (m, 1H), 2.50 (m, 1H), 2.02 (m,1H), 1.43 (m, 15H). MS (M+1): 510.0.

Example 112 tert-Butyl5-(6-(3-(benzyloxy)phenylthio)-1,2,3,4-tetrahydronaphthalen-2-yl)-2,2-dimethyl-1,3-dioxan-5-ylcarbamate

Trifluoromethanesulfonic acid6-(5-tert-butoxycarbonylamino-2,2-dimethyl-1,3-dioxinan-5-yl)-5,6,7,8-tetrahydro-naphthalen-2-ylester (0.310 g, 0.000608 mol), 3-benzyloxy-benzenethiol (0.145 g,0.000669 mol), tris(dibenzylideneacetone)-dipalladium(0) (0.0167 g,0.0000182 mol) and xantphos (0.0211 g, 0.0000365 mol) were dissolved in1,4-dioxane (10 mL, 0.1 mol). The solution was degassed by applying lowvacuum and backfilling with N₂ for 5 times. The pre-degassedN,N-diisopropylethylamine (0.350 mL, 0.00201 mol) in the same way wasadded. The mixture was heated to reflux under an atmosphere of nitrogenfor 14 hours then cooled to r.t.tris(dibenzylideneacetone)dipalladium(0) (0.0501 g, 0.0000548 mol) andxantphos (0.0634 g, 0.000110 mol) was added. The mixture was degassedthree times, heated to reflux for 6 more hours, cooled to r.t. Themixture was filtered, and concentrated to give a crude residue.Chromatography with EtOAc-hexane (0:100 to 20:80) two times gave a solidproduct (243 mg, 43%). ¹H NMR (CHLOROFORM-d) δ: 7.31-7.44 (m, 5H),7.18-7.23 (m, 1H), 7.17 (d, J=2.0 Hz, 2H), 7.04-7.09 (m, 1H), 6.97-7.04(m, 1H), 6.84-6.92 (m, 1H), 6.79-6.84 (m, 1H), 5.02 (s, 2H), 4.86 (s,br, 1H), 3.95-4.05 (m, 4H), 2.77-2.97 (m, 4H), 2.64-2.76 (m, 1H),2.47-2.60 (m, 1H), 2.00-2.14 (m, 1H), 1.51-1.60 (m, 6H), 1.41-1.51 (m,9H). MS (M+Na+): 598.0.

Example 1132-Amino-2-(6-(3-(benzyloxy)phenylthio)-1,2,3,4-tetrahydronaphthalen-2-yl)propane-1,3-diol

tert-Butyl5-(6-(3-(benzyloxy)phenylthio)-1,2,3,4-tetrahydronaphthalen-2-yl)-2,2-dimethyl-1,3-dioxan-5-ylcarbamate(0.240 g, 0.000292 mol) was treated with 6.0 M hydrogen chloride inwater (10 mL, 0.06 mol) in methanol (20 mL, 0.5 mol) at r.t. for 17hours, To the mixture was added ˜2 mL of EtOAc and the mixture wascontinuously stirred at r.t. for 48 hours. The solution was concentratedto dryness and dissolved in methanol, purified by HPLC on a Gilsonsystem. The pure fractions were combined, concentrated and lyophilizedfor 3 days to give a white powder product as TFA salt (0.100 g, yield54.5%). ¹H NMR (MeOD) δ: 7.15-7.30 (m, 5H), 7.04-7.11 (m, 1H), 6.99 (m,3H), 6.65-6.76 (m, 3H), 4.90 (s, 2H), 3.65 (d, J=2.0 Hz, 4H), 2.62-2.83(m, 4H), 2.13-2.27 (m, 1H), 1.88-1.98 (m, 1H), 1.37-1.53 (m, 1H). MS(M+1): 436.0.

Example 114 Sphingosine Kinase Assay

To assay a test compound for its properties as a substrate ofsphingosine kinase 2 (SK2), an assay using recombinantly expressed SK2was used. Briefly, HEK293E cells were transiently transfected withplasmids containing DNA encoding a sphingosine kinase 2 (SK2) (canine,mouse, or human). The cells were cultured in Dulbecco's MinimalEssential Medium (DMEM) containing 0.25 mg/mL G418, 10% fetal calf serum(FCS), and 10 mL/L amphotericin/streptomycin for 48 hours, thenharvested, washed three times in phosphate buffered saline (PBS) andlysed by incubation in lysis buffer (20 mM Tris pH 7.4, 20% glycerol, 1mM β-mercaptoethanol, 1 mM EDTA, 1 mM Na orthovanadate, 40 mMβ-glycerophosphate, 15 mM NaF, 10 mg/mL, leupeptin, 10 mg/mL soybeantrypsin inhibitor, 1 mM PMSF, 0.5 mM 4-deoxyperidoxone, 200 mM KCl, 10mM MgCl₂, for about 30 minutes on ice. The lysate was centrifuged at15,000 rpm for 18 minutes and the cell debris was discarded. The solublefraction was used in the sphingosine kinase reaction. Examples of SPHK2concentrations in the resulting lysates were 8.12 μg/μL (in a canineSPHK2 preparation) and 8.47 μg/μL (in a human SPHK2 preparation).

The SK2 kinase assay was performed in a 200 μL reaction mixturecontaining 20 μm sphingosine (control) or 20 μM test compound (preparedas a 200 μM stock solution containing 0.1% fatty acid-free bovine serumalbumin (BSA)), 38 μL lysate, and 2 μM ATP (freshly prepared). Kinasereactions were incubated at 37° C. for 70 minutes followed by detectionof phosphorylated compound (control or test compound) using UVabsorbance at 282 nm. To further analyze the test compound from thisreaction, test compound kinase reactions were mixed with an equal volumeof acidic acetonitrile and shaken for 10 minutes. The proteinprecipitate was spun down. The supernatant was analyzed on HPLC using aC18 column. The parent and phosphorylated test compound were quantitatedusing area under the curve (AUC) calculation. Test compounds that arephosphorylated in this assay are candidate compounds for use as S1Pmodulators.

Example 115 Lymphopenia Assay

Measurement of circulating lymphocytes: Compounds were dissolved in 30%HPCD. Mice (C57bl/6 male, 6-10 week-old) were administered 0.5 and 5mg/kg of a compound via oral gavage. 30% HPCD was included as a negativecontrol.

Blood was collected from the retro-orbital sinus 5 and 24 hours afterdrug administration under short isoflurane anesthesia. Whole bloodsamples were subjected to hematology analysis. Peripheral lymphocytecounts were determined using an automated analyzer (HEMAVET™ 3700).Three mice were used to assess the lymphocyte depletion activity of eachcompound screened.

Compounds of formula (I) induced full lymphopenia at times as short as 3hours or less to as long as 48 hours or more; for example, 4 to 36hours, or 5 to 24 hours. In some cases, a compound of formula inducedfull lymphopenia at 5 hours and partial lymphopenia at 24 hours. Thedosage required to induce lymphopenia can be in the range of, e.g.,0.001 mg/kg to 100 mg/kg; or 0.01 mg/kg to 10 mg/kg. The dosage can be10 mg/kg or less, such as 5 mg/kg or less, 1 mg/kg or less. or 0.1 mg/kgor less.

Lymphopenia Example # (ED₅₀ mg/kg) 18 0.5-5 mg/kg 21 <0.5 mg/kg 23 <0.5mg/kg 24 >5 mg/kg 26 <0.5 mg/kg 27 >5 mg/kg 38 0.5-5 mg/kg 50 0.5-5mg/kg 61 0.5-5 mg/kg 71 <0.5 mg/kg 71A >5 mg/kg 78 >5 mg/kg 79 >5 mg/kg79A >5 mg/kg 91 0.5-5 mg/kg

These results demonstrated compounds of the invention can inducelymphopenia.

Example 116 Calcium Mobilization

Compounds that were not specific for the S1P₁ receptor, can haveactivity for other S1P receptor subtypes, e.g., S1P₂, S1P₃, S1P₄, orS1P₅, and can cause undesirable side effects. Accordingly, compoundswere tested to identify those that were specific for S1P₁ activity andhad little or no activity, or are antagonists of, S1P₃ activity.Accordingly, the test compounds were tested in a calcium mobilizationassay to determine agonist activity at either the human S1P₁ or humanS1P₃ receptor, and antagonist activity only at the human S1P₃ receptor.The procedure was essentially as described (with modifications describedbelow) in Davis et al. (2005) Journal of Biological Chemistry, vol. 280,pp. 9833-9841, which is incorporated by reference in its entirety.Calcium mobilization assays were performed in recombinant CHEM cellsexpressing human S1P₁ or S1P₃ purchased from Millipore (Billerica,Mass.). To detect free intracellular calcium, S1P₁ or S1P₃ cells wereloaded with FLIPR Calcium 4 dye from Molecular Devices (Sunnyvale,Calif.). Cells were imagined for calcium mobilization using aFLIPR^(TETRA) equipped with a 96-well dispense head.

The data of FIG. 8 show that compounds 21 and 61A are agonists of theS1P₁ receptor, with potencies similar to S1P. The data of FIG. 9 showthat compound 21 has weak micromolar partial S1P₃ agonist activity,whereas no activity was seen for compound 2. S1P was a full S1P₃ agonistin this assay. The data of FIG. 10 show that compounds 21 and 61A do notantagonize the S1P₃ receptor. The data of FIG. 11 show that compounds263 and 269 were S1P₁ agonists, were not S1P₃ agonists, but instead wereS1P₃ antagonists.

Example 117 In Vivo Blood Lymphocyte Depletion

Compounds useful for treating S1P₁-related diseases, such as certainautoimmune diseases, are generally able to sustain lymp-hopenia, e.g.,for at least one day, at least two days, at least three days, or atleast one week, or longer. To further characterize the activity of atest compound, a test compound of formula I or the vehicle wasadministered orally by gavage to rats. Tail blood for hematologicalmonitoring was obtained on day-1 to give the baseline individual values,and at 2, 6, 24, 48 and 72 hours after drug application.

Example 118 In Vivo Screening Assays

Measurement of circulating lymphocytes: Compounds were dissolved in DMSOand further diluted with deionized water. Mice (C57bl/6 male, 6-10week-old) were administered 20 μg of a compound (diluted in 200 μLwater, 4% DMSO) via intra-peritoneal (IP) injection under shortisoflurane anesthesia. 200 μL water, 4% DMSO, and a known S1P agonistwere included as negative controls.

Blood was collected from the retro-orbital sinus 18 hours after drugadministration under short isoflurane anesthesia. Whole blood sampleswere subjected to hematology analysis. Peripheral lymphocyte counts weredetermined using an automated analyzer (HEMAVET™ 3700). Subpopulationsof peripheral blood lymphocytes were stained by fluorochrome-conjugatedspecific antibodies and analyzed using a fluorescent activating cellsorter (FACSCALIBUR™). Two mice were used to assess the lymphocytedepletion activity of each compound screened. This assay indicated thatcompounds of the invention can suppress the level of circulatinglymphocytes.

Example 119 Assessment of Heart Effect

One reported undesirable effect of an S1P agonist can be, e.g.,bradycardia. Assays were conducted to determine the effect of testcompounds on heart function. The effects of compounds on cardiacfunction were monitored using the AnonyMOUSE ECG recording system. ECGswere recorded in conscious mice (C57bl/6 male, 6-10 week-old) before andafter compound administration. 90 μg of compound further diluted in 200μL water and 15% DMSO were injected IP. Four mice were used to assessheart rate effect of each compound. Compounds were found to have littleor no effect on heart rate at therapeutic levels. The abbreviations usedherein have their conventional meaning within the clinical, chemical,and biological arts. In the case of any inconsistencies, the presentdisclosure, including any definitions therein will prevail.

The disclosures of each and every patent, patent application, andpublication cited herein are expressly incorporated herein by referencein their entirety into this disclosure. Illustrative embodiments of thisdisclosure are discussed and reference has been made to possiblevariations within the scope of this disclosure. These and othervariations and modifications in the disclosure will be apparent to thoseskilled in the art without departing from the scope of the disclosure,and it should be understood that this disclosure and the claims shownbelow are not limited to the illustrative embodiments set forth herein.

1. A compound of formula (I):

wherein: A¹ is —C(X¹)═, —N═, —O—, —S—, or a bond; A² is —C(X²)═, —N═,—O—, —S—, or a bond; A³ is —C(X³)(X^(3′))—, —C(X³)═, —NX³—, —N═, —O—, or—S—; A⁴ is —C(X⁴)(X^(4′))—, —C(X⁴)═, —NX⁴—, —N═, —O—, or a bond; A⁵ is—C(X⁵)(X^(5′))—, —C(X⁵)═, —NX⁵—, —N═, —O—, or —S—; A⁶ is —C(X⁶)═, —N═,—O—, —S—, or a bond; provided that A¹, A², A³, A⁴, A⁵ and A⁶ are notsimultaneously —C(X¹)═, —C(X²)═, —C(X³)═, —C(X⁴)═, —C(X⁵)═, and —C(X⁶)═respectively, and provided that the bicyclic ring includes 0-3heteroatoms; and further provided that no more than one of A¹, A², andA⁶ is a bond; X¹ is hydrogen, halo, hydroxy, nitro, cyano, alkyl,haloalkyl, cycloalkyl, halocycloalkyl, alkoxy, haloalkoxy, cycloalkoxy,halocycloalkoxy, acyl, aminoacyl, —NR^(f)R^(g), —N(R^(f))SO₂R^(g),—SO₂R^(f), —SO₂NR^(f)R^(g), —CO₂R^(f), trialkylamino, aryl, orheteroaryl; X² is hydrogen, halo, hydroxy, nitro, cyano, alkyl,haloalkyl, cycloalkyl, halocycloalkyl, alkoxy, haloalkoxy, cycloalkoxy,halocycloalkoxy, acyl, aminoacyl, —NR^(f)R^(g), —N(R^(f))SO₂R^(g),—SO₂R^(f), —SO₂NR^(f)R^(g), —CO₂R^(f), trialkylamino, aryl, orheteroaryl; X³ is hydrogen, halo, hydroxy, nitro, cyano, alkyl,haloalkyl, cycloalkyl, halocycloalkyl, alkoxy, haloalkoxy, cycloalkoxy,halocycloalkoxy, acyl, aminoacyl, —NR^(f)R^(g), —N(R^(f))SO₂R^(g),—SO₂R^(f), —SO₂NR^(f)R^(g), —CO₂R^(f), trialkylamino, aryl, orheteroaryl; X^(3′) is is hydrogen, halo, hydroxy, nitro, cyano, alkyl,haloalkyl, cycloalkyl, halocycloalkyl, alkoxy, haloalkoxy, cycloalkoxy,halocycloalkoxy, acyl, aminoacyl, —NR^(f)R^(g), —N(R^(f))SO₂R^(g),—SO₂R^(f), —SO₂NR^(f)R^(g), —CO₂R^(f), trialkylamino, aryl, orheteroaryl; X⁴ is hydrogen, halo, hydroxy, nitro, cyano, alkyl,haloalkyl, cycloalkyl, halocycloalkyl, alkoxy, haloalkoxy, cycloalkoxy,halocycloalkoxy, acyl, aminoacyl, —NR^(f)R^(g), —N(R^(f))SO₂R^(g),—SO₂R^(f), —SO₂NR^(f)R^(g), —CO₂R^(f), trialkylamino, aryl, orheteroaryl; X^(4′) is is hydrogen, halo, hydroxy, nitro, cyano, alkyl,haloalkyl, cycloalkyl, halocycloalkyl, alkoxy, haloalkoxy, cycloalkoxy,halocycloalkoxy, acyl, aminoacyl, —NR^(f)R^(g), —N(R^(f))SO₂R^(g),—SO₂R^(f), —SO₂NR^(f)R^(g), —CO₂R^(f), trialkylamino, aryl, orheteroaryl; X⁵ is hydrogen, halo, hydroxy, nitro, cyano, alkyl,haloalkyl, cycloalkyl, halocycloalkyl, alkoxy, haloalkoxy, cycloalkoxy,halocycloalkoxy, acyl, aminoacyl, —NR^(f)R^(g), —N(R^(f))SO₂R^(g),—SO₂R^(f), —SO₂NR^(f)R^(g), —CO₂R^(f), trialkylamino, aryl, orheteroaryl; X^(5′) is hydrogen, halo, hydroxy, nitro, cyano, alkyl,haloalkyl, cycloalkyl, halocycloalkyl, alkoxy, haloalkoxy, cycloalkoxy,halocycloalkoxy, acyl, aminoacyl, —NR^(f)R^(g), —N(R^(f))SO₂R^(g),—SO₂R^(f), —SO₂NR^(f)R^(g), —CO₂R^(f), trialkylamino, aryl, orheteroaryl; X⁶ is hydrogen, halo, hydroxy, nitro, cyano, alkyl,haloalkyl, cycloalkyl, halocycloalkyl, alkoxy, haloalkoxy, cycloalkoxy,halocycloalkoxy, acyl, aminoacyl, —NR^(f)R^(g), —N(R^(f))SO₂R^(g),—SO₂R^(f), —SO₂NR^(f)R^(g), —CO₂R^(f), trialkylamino, aryl, orheteroaryl; Y is —OR^(f), —(CR^(f)R^(g))OR^(f), —(CR^(f)R^(g))₂OR^(f),—O—P(O)(OR^(f))OR^(g), —OC(O)R^(c), —C(O)OR^(c),—(CR^(f)R^(g))—P(O)(OR^(f))OR^(g), —(C(OH)R^(f))—P(O)(OR^(f))OR^(g),—S—P(O)(OR^(f))OR^(g), tetrazole, —SO₂NHR^(f), —SO₃, —CONHR^(f),—Si(OH)₂, or —B(OH)₂; W is —CR^(f)R^(g)—, —NR^(f)—, —O—, —S—, —SO—, or—SO₂—; Cy is cycloalkyl, cycloalkenyl, heterocyclyl, aryl, orheteroaryl; wherein Cy is optionally substituted by 1-6 substituentsselected from the group consisting of hydrogen, halo, hydroxy, nitro,cyano, —NR^(f)R^(g), alkyl, haloalkyl, cycloalkyl, cycloalkenyl,cycloalkylalkyl, cycloalkenylalkyl, heterocyclylalkyl, arylalkyl,heteroarylalkyl, alkoxy, haloalkoxy, cycloalkylalkoxy,cycloalkenylalkoxy, heterocyclylalkoxy, aryloxy, arylalkoxy,heteroaryloxy, heteroarylalkoxy, acyl, cycloalkylacyl, cycloalkenylacyl,heterocyclylacyl, arylacyl, heteroarylacyl, thioalkyl, alkenyl, alkynyl,cycloalkenyl, heterocyclyl, aryl, and heteroaryl; L¹ is —CH₂—, —CHF—, or—CF₂—; Z⁴ is hydrogen, halo, alkyl, haloalkyl, alkenyl, haloalkenyl,alkynyl, or —OR^(f); or Z⁴ is —CH₂— bound to the carbon atom to which Yis bound; or L¹, Z⁴, Y, and the atoms to which they are bound form a 4-7membered cycloalkyl group or a 4-7 membered heterocyclyl group having 1or 2 heteroatoms selected from O and N; R^(a) is hydrogen, alkyl,haloalkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, aryl, heteroaryl,or heterocyclyl; wherein each of alkyl, cycloalkyl, cycloalkenyl, aryl,heteroaryl and heterocycle are optionally substituted with 1 to 5substituents independently selected from the group consisting of halo,oxo, —CN, —CHO, —CF₃, —OH, —NO₂, alkyl, —OCF₃, alkoxy, cycloalkoxy,cycloalkenoxy, amino, alkylamino, dialkylamino, acylamino, aminoacyl,alkylsulfonyl, alkylaminosulfonyl, and dialkylaminosulfonyl; R^(b) ishydrogen, alkyl, haloalkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl,aryl, heteroaryl, or heterocyclyl; wherein each of alkyl, cycloalkyl,cycloalkenyl, aryl, heteroaryl and heterocycle are optionallysubstituted with 1 to 5 substituents independently selected from thegroup consisting of halo, oxo, —CN, —CHO, —CF₃, —OH, —NO₂, alkyl, —OCF₃,alkoxy, cycloalkoxy, cycloalkenoxy, amino, alkylamino, dialkylamino,acylamino, aminoacyl, alkylsulfonyl, alkylaminosulfonyl, anddialkylaminosulfonyl; or R^(b) and Z⁴ are taken to together to form—C(O)O— or ═C(R^(f))O—; R^(c) is alkyl, aryl, trifluoromethyl,methylsulfonyl, trifluoromethylsulfonyl, p-tolylsulfonyl, or a groupselected such that −OCOR^(c) is a good leaving group; each R^(f),independently, is hydrogen, alkyl, haloalkyl, alkenyl, alkynyl,cycloalkyl, cycloalkenyl, aryl, heteroaryl, or heterocyclyl; whereineach of alkyl, cycloalkyl, cycloalkenyl, aryl, heteroaryl andheterocycle are optionally substituted with 1 to 5 substituentsindependently selected from the group consisting of halo, oxo, —CN,—CHO, —CF₃, —OH, —NO₂, alkyl, —OCF₃, alkoxy, cycloalkoxy, cycloalkenoxy,amino, alkylamino, dialkylamino, acylamino, aminoacyl, alkylsulfonyl,alkylaminosulfonyl, and dialkylaminosulfonyl; each R^(g), independently,is hydrogen, alkyl, haloalkyl, alkenyl, alkynyl, cycloalkyl,cycloalkenyl, aryl, heteroaryl, or heterocyclyl; wherein each of alkyl,cycloalkyl, cycloalkenyl, aryl, heteroaryl and heterocycle areoptionally substituted with 1 to 5 substituents independently selectedfrom the group consisting of halo, oxo, —CN, —CHO, —CF₃, —OH, —NO₂,alkyl, —OCF₃, alkoxy, cycloalkoxy, cycloalkenoxy, amino, alkylamino,dialkylamino, acylamino, aminoacyl, alkylsulfonyl, alkylaminosulfonyl,and dialkylaminosulfonyl; or a pharmaceutically acceptable salt orprodrug thereof.
 2. The compound of claim 1, wherein W is —O—.
 3. Thecompound of claim 1, wherein R^(a) and R^(b), independently, are each Hor alkyl.
 4. The compound of claim 1, wherein Y is —OR^(f).
 5. Thecompound of claim 1, wherein Y is —OH or —O—P(O)(OR^(f))OR^(g).
 6. Thecompound of claim 1, wherein wherein X⁶ is H, halo, alkyl, cycloalkyl,or haloalkyl.
 7. The compound of claim 1, wherein A³ is —C(X³)H—, A⁴ is—C(X⁴)H—, and A⁵ is is —C(X⁵)H—.
 8. The compound of claim 1, wherein Cyhas the formula:

wherein Z¹ is a bond, —[C(R^(d)R^(e))]_(x)—, —CR^(d)═CR^(e)—, —O—,—NR^(f)—; Z² is a bond, —[C(R^(d)R^(e))]_(y)—, —CR^(d)═CR^(e)—, —O—,—NR^(f)—; Z³ is a bond, —[C(R^(d)R^(e))]_(z)—, —CR^(d)═CR^(e)—, —O—,—NR^(f)—; each of x, y, and z, independently, is 1 to 3; each R^(d),independently, is H, halo, hydroxy, alkyl, haloalkyl, alkenyl, alkoxy,cycloalkyl, —C(O)NR^(f)R^(g), —NR^(f)R^(g), —NR^(f)C(O)R^(g), or—SO₂NR^(f)R^(g); each R^(e), independently, is H, halo, hydroxy, alkyl,haloalkyl, alkenyl, alkoxy, or cycloalkyl, —C(O)NR^(f)R^(g),—NR^(f)R^(g), —NR^(f)C(O)R^(g), or —SO₂NR^(f)R^(g); R^(1a) and R^(1b),independently, are hydrogen, halo, hydroxy, nitro, cyano, —NR^(f)R^(g),alkyl, haloalkyl, cycloalkyl, cycloalkenyl, cycloalkylalkyl,cycloalkenylalkyl, heterocyclylalkyl, arylalkyl, heteroarylalkyl,alkoxy, cycloalkylalkoxy, cycloalkenylalkoxy, heterocyclylalkoxy,arylalkoxy, heteroarylalkoxy, acyl, cycloalkylacyl, cycloalkenylacyl,heterocyclylacyl, arylacyl, heteroarylacyl, thioalkyl, alkenyl, alkynyl,cycloalkenyl, heterocyclyl, aryl, or heteroaryl; or R^(1a) and R^(1b),when taken together, are C₂-C₅ alkylene optionally terminated by orinturrepted by 1 or 2 oxygen atoms, or C₂-C₅ alkenylene optionallyterminated by or inturrepted by 1 or 2 oxygen atoms; R^(2a) and R^(2b),independently, are hydrogen, halo, hydroxy, nitro, cyano, —NR^(f)R^(g),alkyl, haloalkyl, cycloalkyl, cycloalkenyl, cycloalkylalkyl,cycloalkenylalkyl, heterocyclylalkyl, arylalkyl, heteroarylalkyl,alkoxy, cycloalkylalkoxy, cycloalkenylalkoxy, heterocyclylalkoxy,arylalkoxy, heteroarylalkoxy, acyl, cycloalkylacyl, cycloalkenylacyl,heterocyclylacyl, arylacyl, heteroarylacyl, thioalkyl, alkenyl, alkynyl,cycloalkenyl, heterocyclyl, aryl, or heteroaryl; or R^(1a) and R^(2a),when taken together, are C₁-C₅ alkylene optionally terminated by orinturrepted by 1 or 2 oxygen atoms, or C₂-C₅ alkenylene optionallyterminated by or inturrepted by 1 or 2 oxygen atoms; wherein R^(1a),R^(1b), R^(2a), and R^(2b) are each, independently, substituted with 0-5substituents selected from halo, hydroxy, nitro, cyano, —NR^(f)R^(g), or—CO₂R^(f).
 9. The compound of claim 8, wherein R^(1a) and R^(2a) areboth hydrogen.
 10. The compound of claim 8, wherein Z¹ is —CH₂CH₂—. 11.The compound of claim 10, wherein Z² is —CH₂—.
 12. The compound of claim11, wherein Z³ is a bond.
 13. The compound of claim 8, wherein R^(1b) isfluoro, chloro, bromo, iodo, methyl, difluoromethyl, triflurormethyl,ethyl, 1,1-difluoroethyl, propyl, isopropyl, n-butyl, i-butyl, t-butyl,n-pentyl, isopentyl, 1,1-dimethylpropyl, neopentyl, cyclopentyl,n-hexyl, cyclohexyl, methoxy, trifluoromethoxy, ethoxy, n-propoxy,i-propoxy, n-butoxy, i-butoxy, t-butoxy, n-pentyloxy, i-pentyloxy,1,1-dimethylpropoxy, neopentyloxy, cyclopentyloxy, n-hexyloxy, orcyclohexyloxy.
 14. A compound of formula (II):

wherein X¹ is hydrogen, halo, hydroxy, nitro, cyano, alkyl, haloalkyl,cycloalkyl, halocycloalkyl, alkoxy, haloalkoxy, cycloalkoxy,halocycloalkoxy, acyl, aminoacyl, —NR^(f)R^(g), —N(R^(f))SO₂R^(g),—SO₂R^(f), —SO₂NR^(f)R^(g), —CO₂R^(f), trialkylamino, aryl, orheteroaryl; X² is hydrogen, halo, hydroxy, nitro, cyano, alkyl,haloalkyl, cycloalkyl, halocycloalkyl, alkoxy, haloalkoxy, cycloalkoxy,halocycloalkoxy, acyl, aminoacyl, —N(R^(f))SO₂R^(g), —SO₂R^(f),—SO₂NR^(f)R^(g), —CO₂R^(f), trialkylamino, aryl, or heteroaryl; X³ ishydrogen, halo, hydroxy, nitro, cyano, alkyl, haloalkyl, cycloalkyl,halocycloalkyl, alkoxy, haloalkoxy, cycloalkoxy, halocycloalkoxy, acyl,aminoacyl, —NR^(f)R^(g), —N(R^(f))SO₂R^(g), —SO₂R^(f), —SO₂NR^(f)R^(g),—CO₂R^(f), trialkylamino, aryl, or heteroaryl; X^(3′) is is hydrogen,halo, hydroxy, nitro, cyano, alkyl, haloalkyl, cycloalkyl,halocycloalkyl, alkoxy, haloalkoxy, cycloalkoxy, halocycloalkoxy, acyl,aminoacyl, —NR^(f)R^(g), —N(R^(f))SO₂R^(g), —SO₂R^(f), —SO₂NR^(f)R^(g),—CO₂R^(f), trialkylamino, aryl, or heteroaryl; X⁴ is hydrogen, halo,hydroxy, nitro, cyano, alkyl, haloalkyl, cycloalkyl, halocycloalkyl,alkoxy, haloalkoxy, cycloalkoxy, halocycloalkoxy, acyl, aminoacyl,—NR^(f)R^(g), —N(R^(f))SO₂R^(g), —SO₂R^(f), —SO₂NR^(f)R^(g), —CO₂R^(f),trialkylamino, aryl, or heteroaryl; X^(4′) is is hydrogen, halo,hydroxy, nitro, cyano, alkyl, haloalkyl, cycloalkyl, halocycloalkyl,alkoxy, haloalkoxy, cycloalkoxy, halocycloalkoxy, acyl, aminoacyl,—NR^(f)R^(g), —N(R^(f))SO₂R^(g), —SO₂R^(f), —SO₂NR^(f)R^(g), —CO₂R^(f),trialkylamino, aryl, or heteroaryl; X⁵ is hydrogen, halo, hydroxy,nitro, cyano, alkyl, haloalkyl, cycloalkyl, halocycloalkyl, alkoxy,haloalkoxy, cycloalkoxy, halocycloalkoxy, acyl, aminoacyl, —NR^(f)R^(g),—N(R^(f))SO₂R^(g), —SO₂R^(f), —SO₂NR^(f)R^(g), —CO₂R^(f), trialkylamino,aryl, or heteroaryl; X^(5′) is hydrogen, halo, hydroxy, nitro, cyano,alkyl, haloalkyl, cycloalkyl, halocycloalkyl, alkoxy, haloalkoxy,cycloalkoxy, halocycloalkoxy, acyl, aminoacyl, —NR^(f)R^(g),—N(R^(f))SO₂R^(g), —SO₂R^(f), —SO₂NR^(f)R^(g), —CO₂R^(f), trialkylamino,aryl, or heteroaryl; X⁶ is hydrogen, halo, hydroxy, nitro, cyano, alkyl,haloalkyl, cycloalkyl, halocycloalkyl, alkoxy, haloalkoxy, cycloalkoxy,halocycloalkoxy, acyl, aminoacyl, —NR^(f)R^(g), —N(R^(f))SO₂R^(g),—SO₂R^(f), —SO₂NR^(f)R^(g), —CO₂R^(f), trialkylamino, aryl, orheteroaryl; Y is —OR^(f), —(CR^(f)R^(g))OR^(f), —(CR^(f)R^(g))₂OR^(f),—O—P(O)(OR^(f))OR^(g), —OC(O)R^(c), —C(O)OR^(c),—(CR^(f)R^(g))—P(O)(OR^(f))OR^(g), —(C(OH)R^(f))—P(O)(OR^(f))OR^(g),—S—P(O)(OR^(f))OR^(g), tetrazole, —SO₂NHR^(f), —SO₃, —CONHR^(f),—Si(OH)₂, or —B(OH)₂; Z⁴ is hydrogen, halo, alkyl, haloalkyl, alkenyl,haloalkenyl, alkynyl, or —OR^(f); or Z⁴ is —CH₂— bound to the carbonatom to which Y is bound; or Z⁴, Y, and the atoms to which they arebound form a 4-7 membered cycloalkyl group or a 4-7 memberedheterocyclyl group having 1 or 2 heteroatoms selected from O and N;R^(c) is alkyl, aryl, trifluoromethyl, methylsulfonyl,trifluoromethylsulfonyl, p-tolylsulfonyl, or a group selected such that−OCOR^(c) is a good leaving group; each R^(f), independently, ishydrogen, alkyl, haloalkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl,aryl, heteroaryl, or heterocyclyl; wherein each of alkyl, cycloalkyl,cycloalkenyl, aryl, heteroaryl and heterocycle are optionallysubstituted with 1 to 5 substituents independently selected from thegroup consisting of halo, oxo, —CN, —CHO, —CF₃, —OH, —NO₂, alkyl, —OCF₃,alkoxy, cycloalkoxy, cycloalkenoxy, amino, alkylamino, dialkylamino,acylamino, aminoacyl, alkylsulfonyl, alkylaminosulfonyl, anddialkylaminosulfonyl; each R^(g), independently, is hydrogen, alkyl,haloalkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, aryl, heteroaryl,or heterocyclyl; wherein each of alkyl, cycloalkyl, cycloalkenyl, aryl,heteroaryl and heterocycle are optionally substituted with 1 to 5substituents independently selected from the group consisting of halo,oxo, —CN, —CHO, —CF₃, —OH, —NO₂, alkyl, —OCF₃, alkoxy, cycloalkoxy,cycloalkenoxy, amino, alkylamino, dialkylamino, acylamino, aminoacyl,alkylsulfonyl, alkylaminosulfonyl, and dialkylaminosulfonyl; R^(1a) andR^(1b), independently, are hydrogen, halo, hydroxy, nitro, cyano,—NR^(f)R^(g), alkyl, haloalkyl, cycloalkyl, cycloalkenyl,cycloalkylalkyl, cycloalkenylalkyl, heterocyclylalkyl, arylalkyl,heteroarylalkyl, alkoxy, cycloalkylalkoxy, cycloalkenylalkoxy,heterocyclylalkoxy, arylalkoxy, heteroarylalkoxy, acyl, cycloalkylacyl,cycloalkenylacyl, heterocyclylacyl, arylacyl, heteroarylacyl, thioalkyl,alkenyl, alkynyl, cycloalkenyl, heterocyclyl, aryl, or heteroaryl; orR^(1a) and R^(1b), when taken together, are C₂-C₅ alkylene optionallyterminated by or inturrepted by 1 or 2 oxygen atoms, or C₂-C₅ alkenyleneoptionally terminated by or inturrepted by 1 or 2 oxygen atoms; Z¹ is abond, —[C(R^(d)R^(e))]_(x)—, or —CR^(d)═CR^(e)—; Z² is a bond,—[C(R^(d)R^(e))]_(y)—, or —CR^(d)═CR^(e)—; each of x and y,independently, is 1 to 3; each R^(d), independently, is H, halo,hydroxy, alkyl, alkenyl, alkoxy, or cycloalkyl; each R^(e),independently, is H, halo, hydroxy, alkyl, alkenyl, alkoxy, orcycloalkyl; or a pharmaceutically acceptable salt or prodrug thereof.15. The compound of claim 14, wherein Y is —OR^(f).
 16. The compound ofclaim 15, wherein Y is —OH or —O—P(O)(OR^(f))OR^(g).
 17. The compound ofclaim 14, wherein X⁶ is H, halo, alkyl, cycloalkyl, or haloalkyl. 18.The compound of claim 14, wherein Z¹ is —CH₂CH₂—.
 19. The compound ofclaim 18, wherein Z² is —CH₂CH₂—.
 20. The compound of claim 19, whereinR^(1a) is hydrogen, halo, hydroxy, alkyl, haloalkyl, cycloalkyl,cycloalkylalkyl, arylalkyl, alkoxy, cycloalkylalkoxy, arylalkoxy, oraryl.
 21. The compound of claim 14, wherein Y is —OH or —OP(O)(OH)₂; Z⁴is H or —OH; X¹, X², X³, X⁴, and X⁵ are each H; X^(3′), X^(4′), andX^(5′) are each H; and X⁶ is H, halo, alkyl, cycloalkyl, or haloalkyl.22. The compound of claim 14, wherein Z¹ is —(CH₂)_(x)— and Z² is—(CH₂)_(y)—.
 23. The compound of claim 22, wherein R^(1a) is alkyl,haloalkyl, cycloalkyl, aryl, or arylalkoxy.
 24. The compound of claim14, having the formula:


25. A compound of formula (IV):

A³ is —N═, A⁴ is —C(R⁴)═, A⁵ is —C(R⁵)═, and A⁶ is —C(R⁶)═, A³ is—C(R³)═, A⁴ is —N═, A⁵ is —C(R⁵)═, and A⁶ is —C(R⁶)═, A³ is —C(R³)═, A⁴is —C(R⁴)═, A⁵ is —N═, and A⁶ is —C(R⁶)═, A³ is —C(R³)═, A⁴ is —C(R⁴)═,A⁵ is —C(R⁵)═, and A⁶ is —N═, or A³ is —N═, A⁴ is —N═, A⁵ is —C(R⁵)═,and A⁶ is —C(R⁶)═; X¹ is hydrogen, halo, hydroxy, nitro, cyano, alkyl,haloalkyl, cycloalkyl, halocycloalkyl, alkoxy, haloalkoxy, cycloalkoxy,halocycloalkoxy, acyl, aminoacyl, —NR^(f)R^(g), —N(R^(f))SO₂R^(g),—SO₂R^(f), —SO₂NR^(f)R^(g), —CO₂R^(f), trialkylamino, aryl, orheteroaryl; X² is hydrogen, halo, hydroxy, nitro, cyano, alkyl,haloalkyl, cycloalkyl, halocycloalkyl, alkoxy, haloalkoxy, cycloalkoxy,halocycloalkoxy, acyl, aminoacyl, —NR^(f)R^(g), —N(R^(f))SO₂R^(g),—SO₂R^(f), —SO₂NR^(f)R^(g), —CO₂R^(f), trialkylamino, aryl, orheteroaryl; X⁵ is hydrogen, halo, hydroxy, nitro, cyano, alkyl,haloalkyl, cycloalkyl, halocycloalkyl, alkoxy, haloalkoxy, cycloalkoxy,halocycloalkoxy, acyl, aminoacyl, —NR^(f)R^(g), —N(R^(f))SO₂R^(g),—SO₂R^(f), —SO₂NR^(f)R^(g), —CO₂R^(f), trialkylamino, aryl, orheteroaryl; X⁶ is hydrogen, halo, hydroxy, nitro, cyano, alkyl,haloalkyl, cycloalkyl, halocycloalkyl, alkoxy, haloalkoxy, cycloalkoxy,halocycloalkoxy, acyl, aminoacyl, —NR^(f)R^(g), —N(R^(f))SO₂R^(g),—SO₂R^(f), —SO₂NR^(f)R^(g), —CO₂R^(f), trialkylamino, aryl, orheteroaryl; Y is —OR^(f), —(CR^(f)R^(g))OR^(f), —(CR^(f)R^(g))₂OR^(f),—O—P(O)(OR^(f))OR^(g), —OC(O)R^(c), —C(O)OR^(c),—(CR^(f)R^(g))—P(O)(OR^(f))OR^(g), —(C(OH)R^(f))—P(O)(OR^(f))OR^(g),—S—P(O)(OR^(f))OR^(g), tetrazole, —SO₂NHR^(f), —SO₃, —CONHR^(f),—Si(OH)₂, or —B(OH)₂; W is —CR^(f)R^(g)—, —NR^(f)—, —O—, —S—, —SO—, or—SO₂—; Z⁴ is hydrogen, halo, alkyl, haloalkyl, alkenyl, haloalkenyl,alkynyl, or —OR^(f); or Z⁴ is —CH₂— bound to the carbon atom to which Yis bound; or Z⁴, Y, and the atoms to which they are bound form a 4-7membered cycloalkyl group or a 4-7 membered heterocyclyl group having 1or 2 heteroatoms selected from O and N; R^(c) is alkyl, aryl,trifluoromethyl, methylsulfonyl, trifluoromethylsulfonyl,p-tolylsulfonyl, or a group selected such that −OCOR^(c) is a goodleaving group; each R^(f), independently, is hydrogen, alkyl, haloalkyl,alkenyl, alkynyl, cycloalkyl, cycloalkenyl, aryl, heteroaryl, orheterocyclyl; wherein each of alkyl, cycloalkyl, cycloalkenyl, aryl,heteroaryl and heterocycle are optionally substituted with 1 to 5substituents independently selected from the group consisting of halo,oxo, —CN, —CHO, —CF₃, —OH, —NO₂, alkyl, —OCF₃, alkoxy, cycloalkoxy,cycloalkenoxy, amino, alkylamino, dialkylamino, acylamino, aminoacyl,alkylsulfonyl, alkylaminosulfonyl, and dialkylaminosulfonyl; each R^(g),independently, is hydrogen, alkyl, haloalkyl, alkenyl, alkynyl,cycloalkyl, cycloalkenyl, aryl, heteroaryl, or heterocyclyl; whereineach of alkyl, cycloalkyl, cycloalkenyl, aryl, heteroaryl andheterocycle are optionally substituted with 1 to 5 substituentsindependently selected from the group consisting of halo, oxo, —CN,—CHO, —CF₃, —OH, —NO₂, alkyl, —OCF₃, alkoxy, cycloalkoxy, cycloalkenoxy,amino, alkylamino, dialkylamino, acylamino, aminoacyl, alkylsulfonyl,alkylaminosulfonyl, and dialkylaminosulfonyl; R^(1a) and R^(1b),independently, are hydrogen, halo, hydroxy, nitro, cyano, —NR^(f)R^(g),alkyl, haloalkyl, cycloalkyl, cycloalkenyl, cycloalkylalkyl,cycloalkenylalkyl, heterocyclylalkyl, arylalkyl, heteroarylalkyl,alkoxy, cycloalkylalkoxy, cycloalkenylalkoxy, heterocyclylalkoxy,arylalkoxy, heteroarylalkoxy, acyl, cycloalkylacyl, cycloalkenylacyl,heterocyclylacyl, arylacyl, heteroarylacyl, thioalkyl, alkenyl, alkynyl,cycloalkenyl, heterocyclyl, aryl, or heteroaryl; or R^(1a) and R^(1b),when taken together, are C₂-C₅ alkylene optionally terminated by orinturrepted by 1 or 2 oxygen atoms, or C₂-C₅ alkenylene optionallyterminated by or inturrepted by 1 or 2 oxygen atoms; Z¹ is a bond,—[C(R^(d)R^(e))]_(x)—, or —CR^(d)═CR^(e)—; Z² is a bond,—[C(R^(d)R^(e))]_(y)—, or —CR^(d)═CR^(e)—; each of x and yindependently, is 1 to 3; is each R^(d), independently, is hydrogen,halo, hydroxy, alkyl, alkenyl, alkoxy, or cycloalkyl; each R^(e),independently, is hydrogen, halo, hydroxy, alkyl, alkenyl, alkoxy, orcycloalkyl; or a pharmaceutically acceptable salt or prodrug thereof.26. The compound of claim 25, wherein Y is —OR^(f).
 27. The compound ofclaim 26, wherein Y is —OH or —O—P(O)(OR^(f))OR^(g).
 28. The compound ofclaim 25, wherein X⁶ is H, halo, alkyl, cycloalkyl, or haloalkyl. 29.The compound of claim 25, wherein Z¹ is —CH₂CH₂—.
 30. The compound ofclaim 29, wherein Z² is —CH₂CH₂—.
 31. The compound of claim 30, whereinR^(1a) is hydrogen, halo, hydroxy, alkyl, haloalkyl, cycloalkyl,cycloalkylalkyl, arylalkyl, alkoxy, cycloalkylalkoxy, arylalkoxy, oraryl.
 32. A pharmaceutical composition comprising a pharmaceuticallyacceptable carrier and a compound of formula (I):

wherein: A¹ is —C(X¹)═, —N═, —O—, —S—, or a bond; A² is —C(X²)═, —N═,—O—, —S—, or a bond; A³ is —C(X³)(X^(3′))—, —C(X³)═, —NX³—, —N═, —O—, or—S—; A⁴ is —C(X⁴)(X^(4′))—, —C(X⁴)═, —NX⁴—, —N═, —O—, or a bond; A⁵ is—C(X⁵)(X^(5′))—, —C(X⁵)═, —NX⁵—, —N═, —O—, or —S—; A⁶ is —C(X⁶)═, —N═,—O—, —S—, or a bond; provided that A¹, A², A³, A⁴, A⁵ and A⁶ are notsimultaneously —C(X¹)═, —C(X²)═, —C(X³)═, —C(X⁴)═, —C(X⁵)═, and—C(X⁶)═respectively, and provided that the bicyclic ring includes 0-3heteroatoms; and further provided that no more than one of A¹, A², andA⁶ is a bond; X¹ is hydrogen, halo, hydroxy, nitro, cyano, alkyl,haloalkyl, cycloalkyl, halocycloalkyl, alkoxy, haloalkoxy, cycloalkoxy,halocycloalkoxy, acyl, aminoacyl, —NR^(f)R^(g), —N(R^(f))SO₂R^(g),—SO₂R^(f), —SO₂NR^(f)R^(g), —CO₂R^(f), trialkylamino, aryl, orheteroaryl; X² is hydrogen, halo, hydroxy, nitro, cyano, alkyl,haloalkyl, cycloalkyl, halocycloalkyl, alkoxy, haloalkoxy, cycloalkoxy,halocycloalkoxy, acyl, aminoacyl, —NR^(f)R^(g), —N(R^(f))SO₂R^(g),—SO₂R^(f), —SO₂NR^(f)R^(g), —CO₂R^(f), trialkylamino, aryl, orheteroaryl; X³ is hydrogen, halo, hydroxy, nitro, cyano, alkyl,haloalkyl, cycloalkyl, halocycloalkyl, alkoxy, haloalkoxy, cycloalkoxy,halocycloalkoxy, acyl, aminoacyl, —NR^(f)R^(g), —N(R^(f))SO₂R^(g),—SO₂R^(f), —SO₂NR^(f)R^(g), —CO₂R^(f), trialkylamino, aryl, orheteroaryl; X^(3′) is is hydrogen, halo, hydroxy, nitro, cyano, alkyl,haloalkyl, cycloalkyl, halocycloalkyl, alkoxy, haloalkoxy, cycloalkoxy,halocycloalkoxy, acyl, aminoacyl, —NR^(f)R^(g), —N(R^(f))SO₂R^(g),—SO₂R^(f), —SO₂NR^(f)R^(g), —CO₂R^(f), trialkylamino, aryl, orheteroaryl; X⁴ is hydrogen, halo, hydroxy, nitro, cyano, alkyl,haloalkyl, cycloalkyl, halocycloalkyl, alkoxy, haloalkoxy, cycloalkoxy,halocycloalkoxy, acyl, aminoacyl, —NR^(f)R^(g), —N(R^(f))SO₂R^(g),—SO₂R^(f), —SO₂NR^(f)R^(g), —CO₂R^(f), trialkylamino, aryl, orheteroaryl; X^(4′) is is hydrogen, halo, hydroxy, nitro, cyano, alkyl,haloalkyl, cycloalkyl, halocycloalkyl, alkoxy, haloalkoxy, cycloalkoxy,halocycloalkoxy, acyl, aminoacyl, —NR^(f)R^(g), —N(R^(f))SO₂R^(g),—SO₂R^(f), —SO₂NR^(f)R^(g), —CO₂R^(f), trialkylamino, aryl, orheteroaryl; X⁵ is hydrogen, halo, hydroxy, nitro, cyano, alkyl,haloalkyl, cycloalkyl, halocycloalkyl, alkoxy, haloalkoxy, cycloalkoxy,halocycloalkoxy, acyl, aminoacyl, —NR^(f)R^(g), —N(R^(f))SO₂R^(g),—SO₂R^(f), —SO₂NR^(f)R^(g), —CO₂R^(f), trialkylamino, aryl, orheteroaryl; X^(5′) is hydrogen, halo, hydroxy, nitro, cyano, alkyl,haloalkyl, cycloalkyl, halocycloalkyl, alkoxy, haloalkoxy, cycloalkoxy,halocycloalkoxy, acyl, aminoacyl, —NR^(f)R^(g), —N(R^(f))SO₂R^(g),—SO₂R^(f), —SO₂NR^(f)R^(g), —CO₂R^(f), trialkylamino, aryl, orheteroaryl; X⁶ is hydrogen, halo, hydroxy, nitro, cyano, alkyl,haloalkyl, cycloalkyl, halocycloalkyl, alkoxy, haloalkoxy, cycloalkoxy,halocycloalkoxy, acyl, aminoacyl, —NR^(f)R^(g), —N(R^(f))SO₂R^(g),—SO₂R^(f), —SO₂NR^(f)R^(g), —CO₂R^(f), trialkylamino, aryl, orheteroaryl; Y is —OR^(f), —(CR^(f)R^(g))OR^(f), —(CR^(f)R^(g))₂OR^(f),—O—P(O)(OR^(f))OR^(g), —OC(O)R^(c), —C(O)OR^(c),—(CR^(f)R^(g))—P(O)(OR^(f))OR^(g), —(C(OH)R^(f))—P(O)(OR^(f))OR^(g),—S—P(O)(OR^(f))OR^(g), tetrazole, —SO₂NHR^(f), —SO₃, —CONHR^(f),—Si(OH)₂, or —B(OH)₂; W is —CR^(f)R^(g)—, —NR^(f)—, —O—, —S—, —SO—, or—SO₂—; Cy is cycloalkyl, cycloalkenyl, heterocyclyl, aryl, orheteroaryl; wherein Cy is optionally substituted by 1-6 substituentsselected from the group consisting of hydrogen, halo, hydroxy, nitro,cyano, —NR^(f)R^(g), alkyl, haloalkyl, cycloalkyl, cycloalkenyl,cycloalkylalkyl, cycloalkenylalkyl, heterocyclylalkyl, arylalkyl,heteroarylalkyl, alkoxy, haloalkoxy, cycloalkylalkoxy,cycloalkenylalkoxy, heterocyclylalkoxy, aryloxy, arylalkoxy,heteroaryloxy, heteroarylalkoxy, acyl, cycloalkylacyl, cycloalkenylacyl,heterocyclylacyl, arylacyl, heteroarylacyl, thioalkyl, alkenyl, alkynyl,cycloalkenyl, heterocyclyl, aryl, and heteroaryl; L¹ is —CH₂—, —CHF—, or—CF₂—; Z⁴ is hydrogen, halo, alkyl, haloalkyl, alkenyl, haloalkenyl,alkynyl, or —OR^(f); or Z⁴ is —CH₂— bound to the carbon atom to which Yis bound; or L¹, Z⁴, Y, and the atoms to which they are bound form a 4-7membered cycloalkyl group or a 4-7 membered heterocyclyl group having 1or 2 heteroatoms selected from O and N; R^(a) is hydrogen, alkyl,haloalkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, aryl, heteroaryl,or heterocyclyl; wherein each of alkyl, cycloalkyl, cycloalkenyl, aryl,heteroaryl and heterocycle are optionally substituted with 1 to 5substituents independently selected from the group consisting of halo,oxo, —CN, —CHO, —CF₃, —OH, —NO₂, alkyl, —OCF₃, alkoxy, cycloalkoxy,cycloalkenoxy, amino, alkylamino, dialkylamino, acylamino, aminoacyl,alkylsulfonyl, alkylaminosulfonyl, and dialkylaminosulfonyl; R^(b) ishydrogen, alkyl, haloalkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl,aryl, heteroaryl, or heterocyclyl; wherein each of alkyl, cycloalkyl,cycloalkenyl, aryl, heteroaryl and heterocycle are optionallysubstituted with 1 to 5 substituents independently selected from thegroup consisting of halo, oxo, —CN, —CHO, —CF₃, —OH, —NO₂, alkyl, —OCF₃,alkoxy, cycloalkoxy, cycloalkenoxy, amino, alkylamino, dialkylamino,acylamino, aminoacyl, alkylsulfonyl, alkylaminosulfonyl, anddialkylaminosulfonyl; or R^(b) and Z⁴ are taken to together to form—C(O)O— or ═C(R^(f))O—; R^(c) is alkyl, aryl, trifluoromethyl,methylsulfonyl, trifluoromethylsulfonyl, p-tolylsulfonyl, or a groupselected such that −OCOR^(c) is a good leaving group; each R^(f),independently, is hydrogen, alkyl, haloalkyl, alkenyl, alkynyl,cycloalkyl, cycloalkenyl, aryl, heteroaryl, or heterocyclyl; whereineach of alkyl, cycloalkyl, cycloalkenyl, aryl, heteroaryl andheterocycle are optionally substituted with 1 to 5 substituentsindependently selected from the group consisting of halo, oxo, —CN,—CHO, —CF₃, —OH, —NO₂, alkyl, —OCF₃, alkoxy, cycloalkoxy, cycloalkenoxy,amino, alkylamino, dialkylamino, acylamino, aminoacyl, alkylsulfonyl,alkylaminosulfonyl, and dialkylaminosulfonyl; each R^(g), independently,is hydrogen, alkyl, haloalkyl, alkenyl, alkynyl, cycloalkyl,cycloalkenyl, aryl, heteroaryl, or heterocyclyl; wherein each of alkyl,cycloalkyl, cycloalkenyl, aryl, heteroaryl and heterocycle areoptionally substituted with 1 to 5 substituents independently selectedfrom the group consisting of halo, oxo, —CN, —CHO, —CF₃, —OH, —NO₂,alkyl, —OCF₃, alkoxy, cycloalkoxy, cycloalkenoxy, amino, alkylamino,dialkylamino, acylamino, aminoacyl, alkylsulfonyl, alkylaminosulfonyl,and dialkylaminosulfonyl; or a pharmaceutically acceptable salt orprodrug thereof.
 33. A method of making a compound of formula (I)comprising contacting a compound of formula (III):

wherein A¹ is —C(X¹)═, —N═, —O—, —S—, or a bond; A² is —C(X²)═, —N═,—O—, —S—, or a bond; A³ is —C(X³)(X^(3′))—, —C(X³)═, —NX³—, —N═, —O—, or—S—; A⁴ is —C(X⁴)(X^(4′))—, —C(X⁴)═, —NX⁴—, —N═, —O—, or a bond; A⁵ is—C(X⁵)(X^(5′))—, —C(X⁵)═, —NX⁵—, —N═, —O—, or —S—; A⁶ is —C(X⁶)═, —N═,—O—, —S—, or a bond; provided that A¹, A², A³, A⁴, A⁵ and A⁶ are notsimultaneously —C(X¹)═, —C(X²)═, —C(X³)═, —C(X⁴)═, —C(X⁵)═, and—C(X⁶)═respectively, and provided that the bicyclic ring includes 0-3heteroatoms; and further provided that no more than one of A¹, A², andA⁶ is a bond; X¹ is hydrogen, halo, hydroxy, nitro, cyano, alkyl,haloalkyl, cycloalkyl, halocycloalkyl, alkoxy, haloalkoxy, cycloalkoxy,halocycloalkoxy, acyl, aminoacyl, —NR^(f)R^(g), —N(R^(f))SO₂R^(g),—SO₂R^(f), —SO₂NR^(f)R^(g), —CO₂R^(f), trialkylamino, aryl, orheteroaryl; X² is hydrogen, halo, hydroxy, nitro, cyano, alkyl,haloalkyl, cycloalkyl, halocycloalkyl, alkoxy, haloalkoxy, cycloalkoxy,halocycloalkoxy, acyl, aminoacyl, —NR^(f)R^(g), —N(R^(f))SO₂R^(g),—SO₂R^(f), —SO₂NR^(f)R^(g), —CO₂R^(f), trialkylamino, aryl, orheteroaryl; X³ is hydrogen, halo, hydroxy, nitro, cyano, alkyl,haloalkyl, cycloalkyl, halocycloalkyl, alkoxy, haloalkoxy, cycloalkoxy,halocycloalkoxy, acyl, aminoacyl, —NR^(f)R^(g), —N(R^(f))SO₂R^(g),—SO₂R^(f), —SO₂NR^(f)R^(g), —CO₂R^(f), trialkylamino, aryl, orheteroaryl; X^(3′) is is hydrogen, halo, hydroxy, nitro, cyano, alkyl,haloalkyl, cycloalkyl, halocycloalkyl, alkoxy, haloalkoxy, cycloalkoxy,halocycloalkoxy, acyl, aminoacyl, —NR^(f)R^(g), —N(R^(f))SO₂R^(g),—SO₂R^(f), —SO₂NR^(f)R^(g), —CO₂R^(f), trialkylamino, aryl, orheteroaryl; X⁴ is hydrogen, halo, hydroxy, nitro, cyano, alkyl,haloalkyl, cycloalkyl, halocycloalkyl, alkoxy, haloalkoxy, cycloalkoxy,halocycloalkoxy, acyl, aminoacyl, —NR^(f)R^(g), —N(R^(f))SO₂R^(g),—SO₂R^(f), —SO₂NR^(f)R^(g), —CO₂R^(f), trialkylamino, aryl, orheteroaryl; X^(4′) is is hydrogen, halo, hydroxy, nitro, cyano, alkyl,haloalkyl, cycloalkyl, halocycloalkyl, alkoxy, haloalkoxy, cycloalkoxy,halocycloalkoxy, acyl, aminoacyl, —NR^(f)R^(g), —N(R^(f))SO₂R^(g),—SO₂R^(f), —SO₂NR^(f)R^(g), —CO₂R^(f), trialkylamino, aryl, orheteroaryl; X⁵ is hydrogen, halo, hydroxy, nitro, cyano, alkyl,haloalkyl, cycloalkyl, halocycloalkyl, alkoxy, haloalkoxy, cycloalkoxy,halocycloalkoxy, acyl, aminoacyl, —NR^(f)R^(g), —N(R^(f))SO₂R^(g),—SO₂R^(f), —SO₂NR^(f)R^(g), —CO₂R^(f), trialkylamino, aryl, orheteroaryl; X^(5′) is hydrogen, halo, hydroxy, nitro, cyano, alkyl,haloalkyl, cycloalkyl, halocycloalkyl, alkoxy, haloalkoxy, cycloalkoxy,halocycloalkoxy, acyl, aminoacyl, —NR^(f)R^(g), —N(R^(f))SO₂R^(g),—SO₂R^(f), —SO₂NR^(f)R^(g), —CO₂R^(f), trialkylamino, aryl, orheteroaryl; X⁶ is hydrogen, halo, hydroxy, nitro, cyano, alkyl,haloalkyl, cycloalkyl, halocycloalkyl, alkoxy, haloalkoxy, cycloalkoxy,halocycloalkoxy, acyl, aminoacyl, —NR^(f)R^(g), —N(R^(f))SO₂R^(g),—SO₂R^(f), —SO₂NR^(f)R^(g), —CO₂R^(f), trialkylamino, aryl, orheteroaryl; and R³ has the formula:

wherein Z⁴ is H or —OR^(f); and Pg is an amino protecting group; with acompound having the formula:Cy-OH; wherein Cy is cycloalkyl, cycloalkenyl, heterocyclyl, aryl, orheteroaryl; wherein Cy is optionally substituted by 1-6 substituentsselected from the group consisting of hydrogen, halo, hydroxy, nitro,cyano, —NR^(f)R^(g), alkyl, haloalkyl, cycloalkyl, cycloalkenyl,cycloalkylalkyl, cycloalkenylalkyl, heterocyclylalkyl, arylalkyl,heteroarylalkyl, alkoxy, haloalkoxy, cycloalkylalkoxy,cycloalkenylalkoxy, heterocyclylalkoxy, aryloxy, arylalkoxy,heteroaryloxy, heteroarylalkoxy, acyl, cycloalkylacyl, cycloalkenylacyl,heterocyclylacyl, arylacyl, heteroarylacyl, thioalkyl, alkenyl, alkynyl,cycloalkenyl, heterocyclyl, aryl, and heteroaryl; R^(a) is hydrogen,alkyl, haloalkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, aryl,heteroaryl, or heterocyclyl; wherein each of alkyl, cycloalkyl,cycloalkenyl, aryl, heteroaryl and heterocycle are optionallysubstituted with 1 to 5 substituents independently selected from thegroup consisting of halo, oxo, —CN, —CHO, —CF₃, —OH, —NO₂, alkyl, —OCF₃,alkoxy, cycloalkoxy, cycloalkenoxy, amino, alkylamino, dialkylamino,acylamino, aminoacyl, alkylsulfonyl, alkylaminosulfonyl, anddialkylaminosulfonyl; each R^(f), independently, is hydrogen, alkyl,haloalkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, aryl, heteroaryl,or heterocyclyl; wherein each of alkyl, cycloalkyl, cycloalkenyl, aryl,heteroaryl and heterocycle are optionally substituted with 1 to 5substituents independently selected from the group consisting of halo,oxo, —CN, —CHO, —CF₃, —OH, —NO₂, alkyl, —OCF₃, alkoxy, cycloalkoxy,cycloalkenoxy, amino, alkylamino, dialkylamino, acylamino, aminoacyl,alkylsulfonyl, alkylaminosulfonyl, and dialkylaminosulfonyl; each R^(g),independently, is hydrogen, alkyl, haloalkyl, alkenyl, alkynyl,cycloalkyl, cycloalkenyl, aryl, heteroaryl, or heterocyclyl; whereineach of alkyl, cycloalkyl, cycloalkenyl, aryl, heteroaryl andheterocycle are optionally substituted with 1 to 5 substituentsindependently selected from the group consisting of halo, oxo, —CN,—CHO, —CF₃, —OH, —NO₂, alkyl, —OCF₃, alkoxy, cycloalkoxy, cycloalkenoxy,amino, alkylamino, dialkylamino, acylamino, aminoacyl, alkylsulfonyl,alkylaminosulfonyl, and dialkylaminosulfonyl.
 34. A method forprevention or treatment of a pathological condition or symptom in amammal, wherein the activity of sphingosine 1-phosphate receptors isimplicated and agonism of such activity is desired, comprisingadministering to said mammal an effective amount of a compound ofclaim
 1. 35. The method of claim 34, wherein the pathological conditionis neuropathic pain.
 36. The method of claim 34, wherein thepathological condition is an autoimmune disease.
 37. The method of claim36, further comprising administering to said mammal an effective amountof a drug selected from the group consisting of: corticosteroids,bronchodilators, antiasthmatics, antiinflammatories, antirheumatics,immunosuppressants, antimetabolites, immunomodulators, antipsoriatics,and antidiabetics.
 38. The method of claim 36, wherein the autoimmunedisease is uveitis, type I s diabetes, rheumatoid arthritis,inflammatory bowel diseases, lupus, asthma, psoriasis, or multiplesclerosis.
 39. The method of claim 37, wherein the autoimmune disease ismultiple sclerosis.
 40. The method of claim 39, wherein the preventionor treatment of the pathological condition is altering lymphocytetrafficking.
 41. The method of claim 40, wherein altering lymphocytetrafficking provides prolonged allograft survival.
 42. The method ofclaim 41, wherein the allograft is for transplantation.
 43. A method forprevention or treatment of a pathological condition or symptom in amammal, wherein the activity S1P lyase implicated and inhibition of theS1P lyase is desired, comprising administering to said mammal aneffective amount of a compound of claim
 1. 44. An assay, comprising:transfecting HEK293 cells with a plasmid encoding sphingosine kinase 2;obtaining a soluble cell lysate including sphingosine kinase 2;contacting the soluble cell lysate with ATP and a test compound; anddetermining whether the test compound is phosphorylated.